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 a'$H| J  2 H+	
/
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^

J`p.i e(  l7		
dRt[Rb]= 2  RRj
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Sy..z 7dtb%7-    !8FiN	gq5[zFe9y*V
 l `n	DNc}N{    1kK6i	


-B:<2  u>h5m	Rpy-EAdyc8	$6a   Nstst#0189l				uvwx$,MNO@4YZcwx";<H^<	
#45mnC u$2{l:XH	P		"%Pu~5[/#=I_=o'   Oy,b&t+L			
 m_qoO# A%	Mm;	##vN_PYl06f6 v3#;U	I	Q		r]6\0i 6>}y) 6|4Y
_B$J`v>D+2 {;I}x
9YiQM&Q	,jp JU\c&,A
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tn`	g ( x Pz'u
#N

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_"H1 hO
a BRq(			f$`-f~7hg" R <	

$"0	OkQ( G'&9d3njo	
R

5k7w(F3Zmi kR		M tj7"@P: ?w4$rGVcJ		v
	s*!] -Lg4B+ V<PE	4	:`^G1J? 	z| b0	mz7y~|sk 
ICZ	 	c


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ntU`<bi|2mISA39Ka< .X}6df %D"  # y
[]b[Nt>r^kq. p<H~>			Y
 d:FZjR''>-{h#  ,z^$&8RV<`
p!  $|!NT{JGzj. n | 5]0'-		=}  KEB	#JYbL Udz	
-|"} ?QVtlCsVsu = `  
'u	
>o$
u!  dy.<^03N	

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+j4h0.E-\q: y  #O
:JnV	1_[x  {vI[D QZnSWY"  K ]o$sd9/a  Er f	s
3q{jm0~   ,u			.}o~Q  d'gs P C	`32s@ GeGZjV$ 8 o     ci-?&EA /ImN) + r ;*OS	8
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1%B` G-T%;E6XlEG" S =	
e
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Ici#
ZRT#  w4S


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6X058xqd`I$ \  'jk
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M-|M? x+z%sWK		Q
!ot+DL^.!733 @~5LH|w

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sTC  t Q N8=QnQ
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		5	?		
;alHR@R)28@CyU'   "  oSF			%U_
q]zm { 4,{k<B|U _  x&/t1	=Xqptl Ymg
*3VX^  c\nc8prz/}   b,7
@2?M7  i  )p0?ZDg	8	


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r
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$.r5? (+HBx @FTl  l%W)jAT.K    (ao+>yf7	




|v?Va.n}W3VJjB:@Xb>FLB 86Q	A	
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opKLMuF
-=>cdjklm*+,_nopU=?FQL=KIJ; 04ax	d			

,1 BNe[`xF#+84O#CK  		2,9M&&4/ }/h Y!5b	

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egF\.y* f="c<&		"D[k`Sg (b%:+" S~p		!G&|$8i   ;c	


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:$		9;%fY	   AUZ	/Vo  +D}O){k%JEM9# %\K
.'RL "|Hf		hLR 6Y^
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>1nEKx  .1-		

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.Zs_ +~`W WJ	 .A+1- V	

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m[!;Dt\*t%   @4f1EyO&5@v6c# % #t+wL(QcDuWXh) > 1}	$

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LU^bx0pz@#*# G a a (
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
tbvG]n"l  z	8hqkC/  T["1@
,}fq gFg'			!^,2e >Uqb3	u
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Z#E\aZ&-;=sog  T	"K `			vo#k'%j ( 5  tHm9  Jq+t	r
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iJ]U.l]r
C?#  <-^Mw		i
O*019_( W


&,Tnhr5TXi{>R/ SD#   v
kBGO'pF6;_e  
`g v ${~	CK6 `m	P
TQ7w2p\>y ^ z  3w.s+;bUo{Wq
]#  h  h	

69\ 3   |)K
\-kB '.nw1	


0J"El,)^*7&>EWrINR&JB   |)g? 
(^EHJLTd Ulm	vYZ($ BRo;%	f<T9#7x
!   =9		:gXJZ C[	&QU8   %?-U^	
FPp $Bi	p *e	^ V 
y*j0U&  "L*sJ[	%P0t :?;Wb%  P|} z&K@T]rTw%Y"F[)  !g|p: ,E~LTF}	t
l4e,~rn1N ' 4  s*s	jk% ]VLv	N/SNI5	R^ u v-lr	~BTpv="  &f	

/!]+(6Mj T<	P8XI  >#~ITh		
i)>T] S\$i! e	O7MSu] }g	(htZ[\QRS  -  qGACNNNNNNGTC,CCAGGCCTGG,GATCCGGATC,CCWGGTACCWGG,CCANNNNNTGG,CCAGGNNNTGG,CCANNNCCTGG,CCAGGNCCTGG,GCACNNNAACGTTNNNGTGC,CACNNNGTG,CCNNNNNNNGG,AACNAACRYGTGCGTGC,GCACGCACRYGTTNGTT,GCACNAACRYGTTNGTGC,AACGCACRYGTGCGTT,GACNNNNNGTC,GAANNNNNNNTTGG,CCAANNNNNNNTTC,CACNNNNGTG,AACACNNNNGTGC,GCACNNNNGTGTT,GCANNNNNNTGC,GAACNNNNNNTCC,GGANNNNNNGTTC,CAGNNNCTG,CCAGGNNCTG,CAGNNCCTGG,CCWGGGCCC,GGGCCCWGG,CCWGGGCCCWGG,AACNNNNNNGTGCACNNNNNNGTT,GACNNNGTC,GAANNNNTTC,CCWGGTACC,GGTACCWGG,CCWGGWCCWGG,ACNNNNGTAYC,GRTACNNNNGT,CCWGGYRCCWGG,GAAGNNNNNNTAC,GTANNNNNNCTTC,AACGGCNNNGTGC,GCACNNNGCCGTT,GCACGGCNNNGTT,AACNNNGCCGTGC,CGANNNNNNTGC,GCANNNNNNTCG,CGATCNNNNTGC,GCANNNNGATCG,TGANNNNNNTCA,TGATCNNNNTCA,TGANNNNGATCA,TGATCNNGATCA,AACCTGCNNGTGC,GCACNNGCAGGTT,GCACCTGCNNGTT,AACNNGCAGGTGC,GCCNNNNNGGC,AACNNNNNNGTGCMC,GKGCACNNNNNNGTT,CCWGGNCCWGG,AACTGGGNNGTGC,GCACNNCCCAGTT,GCACTGGGNNGTT,AACNNCCCAGTGC,GACNNNNGTC,GAGNNNNNCTC,CCWGGTCTC,GAGACCWGG,GATNNNNATC,GATCNNNATC,GATNNNGATC,GATCNNGATC,CCWGGCGCCWGG,ACNNNNNCTCC,GGAGNNNNNGT,AACNNNNNNGTGCAG,CTGCACNNNNNNGTT,GATCATGATC,CCWGGNNCCWGG,AACTGGNNNGTGC,GCACNNNCCAGTT,GCACTGGNNNGTT,AACNNNCCAGTGC,AACNAACCGGTGCGTGC,GCACGCACCGGTTNGTT,GCACNAACCGGTTNGTGC,AACGCACCGGTGCGTT,GCANNNNNTGC,GCACNNNNNNGTTCGAACNNNNNNGTGC,CCTNNNNNAGG,GCNNNNNNNGC,CCANNNNNNTGG,CCAGGNNCCTGG,CCTGGCCAGG,CAANNNNNGTGG,CCACNNNNNTTG,TTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAA,GCACNNNNNNGTTTAAACNNNNNNGTGC,CCAGGNCCY,RGGNCCTGG,CCWGGGNCCTGG,CCAGGNCCCWGG,AAGNNNNNCTT,GAYNNNNNVTC,GAGNNNNNATC,GAGNNNNNGTC,GATCNNNNVTC,GABNNNNGATC,GAYNNNNGATC,GATCNNNNRTC,GATCNNNGATC,GTYAACNNNNNNGTGC,GCACNNNNNNGTTRAC,GCACNNNNNNGTTAACNNNNNNGTGC,GATCNGATC,TAACTATAACGGTCCTAAGGTAGCGA,TCGCTACCTTAGGACCGTTATAGTTA,CTCTCTTAAGGTAGC,GCTACCTTAAGAGAG,TAGGGATAACAGGGTAAT,ATTACCCTGTTATCCCTA,GTTAACNNNNNNGTGC,GCACNNNNNNGTTAAC,AACNAACGCGTGCGTGC,GCACGCACGCGTTNGTT,GCACNAACGCGTTNGTGC,AACGCACGCGTGCGTT,TCCAACNNNNNNGTGC,GCACNNNNNNGTTGGA,CAYNNNNRTG,AACAYNNNNGTGC,GCACNNNNRTGTT,GTTTAAACNNNNNNGTGC,GCACNNNNNNGTTTAAAC,CCWGGCGCC,GGCGCCWGG,CCWGGNNCC,GGNNCCWGG,GATCGCGATC,AACNAACATGTGCGTGC,GCACGCACATGTTNGTT,GCACNAACATGTTNGTGC,AACGCACATGTGCGTT,TGGCAAACAGCTATTATGGGTATTATGGGT,ACCCATAATACCCATAATAGCTGTTTGCCA,ATCTATGTCGGGTGCGGAGAAAGAGGTAATGAAATGG,CCATTTCATTACCTCTTTCTCCGCACCCGACATAGAT,AACNNGCACGTGCNNGTT,GAACNNNNNCTC,GAGNNNNNGTTC,CCAGGWCCY,RGGWCCTGG,CCWGGGWCCTGG,CCAGGWCCCWGG,CCAGGWCCTGG,TTATAACNNNNNNGTGC,GCACNNNNNNGTTATAA,GCACNNNNNNGTTATAACNNNNNNGTGC,GAACNNNNNNTAC,GTANNNNNNGTTC,GGCCNNNNNGGCC,AACNNACTAGTGC,GCACTAGTNNGTT,AACTAGTNNGTGC,GCACNNACTAGTT,AACNAACTAGTGCGTGC,GCACGCACTAGTTNGTT,GCACNAACTAGTTNGTGC,AACGCACTAGTGCGTT,AACNNNAACGTGCNNGTGC,GCACNNGCACGTTNNNGTT,GCACCCANNNGTT,AACNNNTGGGTGC,TTAACNNNNNNGTGC,GCACNNNNNNGTTAA,CACNNNNNNTCC,GGANNNNNNGTG,CACNNNNGATCC,GGATCNNNNGTG,GATCTAGATC,CCANNNNNNNNNTGG,CCAGGCCT,AGGCCTGG,CCTCGAGG,GATCCGGA,TCCGGATC,GGCGCGCC,GCGATCGC,CCWGGWCC,GGWCCWGG,GATCGATC,GATCATGA,TCATGATC,GCGGCCGC,CCTGGCCR,YGGCCAGG,CCWGGNCC,GGNCCWGG,GGCCGGCC,RTGCGCAY,GATCNNGA,TCNNGATC,CGCGCGCG,CCTGGCCA,TGGCCAGG,CGCCGGCG,GTTTAAAC,GATCGCGA,TCGCGATC,TTAATTAA,VCTCGAGB,CCTGCAGG,CRCCGGYG,CGTCGACG,ATTTAAAT,GCCCGGGC,GATCTAGA,TCTAGATC,CACCTGC,GCAGGTG,CCTNAGG,GATCGAT,ATCGATC,CCTCAGC,GCTGAGG,GCTNAGC,CCTNAGC,GCTNAGG,GCTCTTC,GAAGAGC,GGTNACC,CGGWCCG,ACCWGGT,RGGNCCY,CCTGGAG,CTCCAGG,GGTGATC,GATCACC,GATCNGA,TCNGATC,GGGWCCC,GAAGATC,GATCTTC,CCCWGGG,TCCNGGA,TCCWGGA,RGGWCCY,TTATAA,AGGCCT,GACGTC,GTMKAC,TCCGGA,TGCGCA,ACCTGC,GCAGGT,GGTACC,AGTACT,GGYRCC,CCGCTC,GAGCGG,AACGTT,YGGCCR,RAATTY,CTGAAG,CTTCAG,CACGTG,GRCGYC,AGCGCT,CTTAAG,ACRYGT,ACCGGT,ACTAGT,CACGTC,GACGTG,GWGCWC,GTGCAC,CYCGRG,GGGCCC,ATTAAT,CCTAGG,TTCGAA,GCTAGC,GKGCMC,TGGCCA,GGATCC,GRGCYC,ATCGAT,CACGAG,CTCGTG,GGCGCC,GAAGAC,GTCTTC,GCATGC,GTATCC,GGATAC,TGATCA,ACTGGG,CCCAGT,CTRYAG,RGCGCY,AGATCT,GGNNCC,CTGGAG,CTCCAG,CTTGAG,CTCAAG,CGATCG,GGTCTC,GAGACC,YACGTR,CCNNGG,GAATGC,GCATTC,WCCGGW,GCAATG,CATTGC,RCCGGY,GCGCGC,GAGGAG,CTCCTC,CGGCCG,CCCAGC,GCTGGG,GTGCAG,CTGCAC,CGRYCG,CGTACG,CGTCTC,GAGACG,CCATGG,TCGCGA,GDGCHC,TGTACA,TCATGA,ACATGT,GTATAC,CCWWGG,CTCTTC,GAAGAG,GCNNGC,CCRYGG,GTTAAC,RCATGY,TACGTA,RGATCY,GCGATG,CATCGC,GCAGTG,CACTGC,CCCGGG,CCGCGG,TTTAAA,GGCGGA,TCCGCC,GAGCTC,GATATC,CTGRAG,CTYCAG,CAGCAG,CTGCTG,GAATTC,ATGCAT,CATATG,GTYRAC,AAGCTT,GTNNAC,TCNNGA,CAATTG,ACGCGT,TCCRAC,GTYGGA,GCCGGC,CMGCKG,GCCGAG,CTCGGC,CTCGAG,CTGCAG,CAGCTG,GTCGAC,CTYRAG,AATATT,GATCGA,TCGATC,GACCGA,CACCCA,TCGGTC,TGGGTG,WGTACW,TARCCA,TGGYTA,TCTAGA,GGATC,GATCC,CCWGG,GTCTC,GAGAC,GCWGC,GGNCC,CCSGG,GGTGA,TCACC,GGWCC,GCAGC,GCTGC,CCATC,GATGG,ACGGC,GCCGT,GCNGC,CCNGG,GCATC,GATGC,ACTGG,CCAGT,GGATG,CATCC,CTCAG,CTGAG,GGGAC,GTCCC,ACNGT,CTNAG,CAGTG,CACTG,GACGC,GCGTC,CCCGC,GCGGG,GANTC,CGWCG,TCNGA,CCTTC,GAAGG,GTNAC,GAAGA,TCTTC,GAGTC,GACTC,GTSAC,GAWTC,GCSGC,CASTG,ATGAA,TTCAT,ACGGA,TCCGT,CGCG,CCGC,GCGG,GTAC,AGCT,GCGC,CTAG,GATC,GGCC,CCGG,CATG,RGCY,YATR,ACGT,TGCA,AATT,CCTC,GAGG,TTAA,ASST,TCGA                                                                                                                                                                                                                                                                                                                                                            +                                                     
                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                      	                                                                                                                                 ,               
                        	                                              !                                                                                
   	                  	                              $            
   
      	                                          	                                                      &   '   
   (   $         '            {         )         2   a                                                                                                                                                                       +                         
                                                                                                                                                                                                                                                                                                                              	                                                 ,     
          	                !                              
 	      	          $    
 
  	              	                  & ' 
 ( $   '    {   )   2 a       8  T      V
tu I	*r)*p0"/1 #q
j^`OR4cccc{XvS 'OR[dS<;4=)B
 5)I&Zc eDkDN	 y	+@(y	( r*	
wQm{3^#I_ ry	C$|o
oHo
Hlz(co
Hb	rU.	q$!^ sT>ruCw$ArW	,S 6x<d,}|S\	Xoj$]EO><_		o
oHof&		+=J4 $QF
yKmqu2EOSDX\4=ZB
	,"xh6t9	mVD .  
I kO6dn
k(o

FH g  f"\cK&			!eU+TJ A$EO .
<:Z\blZ\ l6x	0>[e
o1dixb6 #zi|&T]wI	)	#		$	d- 0  	9F&o^d<'R	?Zo"FJ >~T	
		:d~p|'J- 0  	9F&o^dLF,~rn]	| -} x )- 0  !p)r	T
vOX9F&o^:d >~T|	
&T]wI	)xW
Y<f&c2i

4j'Eh) 6 ($h-\q } %"F

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gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7354-7370" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="peroxidase/catalase" directionality="1" translationMW="21996.53" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="7349" maxFusedRunOn="7349"><Segment range="6747-7327" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="3" name="FTL_1504" directionality="1" type="gene" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6747-7327" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="Francisella tularensis subsp. holarctica" type="source" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7327" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="1" name="probably start site" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6708-6710" color="#a6acb3" type="standard"/></Feature><Feature recentID="0" name="mod rbs" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6697-6702" color="#a6acb3" type="standard"/></Feature></Features>
  !<?xml version="1.0"?><Primers nextValidID="13"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite></Primer><Primer recentID="1" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="2" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="3" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="4" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="5" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="6" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="7" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="8" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="9" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="10" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="11" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7381-7400" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="7381-7400" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7381-7400" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="7381-7400" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="12" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="17:54:14">2019.5.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAE26ieeVMoYze*aE*Eۙznvj꒫xggYV^{UZ^eUkUD먵ZwnyzeVVjyyUf[ze>WeJ5fQ&{^LAZ}TU^:X9XgqU[}2FhjVSeSU^Fx]:UQnKu>8jZ`6f$A]PY8=buMC^Y⭂hN	⮤[X>d
$Ex5a^ yDeXD'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ       -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features recycledIDs="6,7,8,0" nextValidID="22"><Feature recentID="5" name="rbs" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6697-6702" color="#a6acb3" type="standard"/></Feature><Feature recentID="4" name="probably start site" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6708-6710" color="#a6acb3" type="standard"/></Feature><Feature recentID="3" name="Francisella tularensis subsp. holarctica" type="source" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7327" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="2" name="FTL_1504" directionality="1" type="gene" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6747-7327" color="#00e600" type="standard"/></Feature><Feature recentID="1" name="peroxidase/catalase" directionality="1" translationMW="21996.53" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="7349" maxFusedRunOn="7349"><Segment range="6747-7327" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="21" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7354-7370" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="16" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="15" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="12" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="11" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7762-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="10" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7328-7332" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="9" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-8056" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  !<?xml version="1.0"?><Primers recycledIDs="5,4,3,0,2,1" nextValidID="19"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="6" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite></Primer><Primer recentID="7" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="9" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="10" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="11" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="12" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="13" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="16" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="17" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7381-7400" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="7381-7400" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7381-7400" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="7381-7400" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="18" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="16:29:3">2019.5.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  AI   '             .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
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ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NA'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  )<?xml version="1.0"?><Features recycledIDs="1,2,3,4,5,6,7,8,0" nextValidID="22"><Feature recentID="21" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6134-6150" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="16" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="15" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="12" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="11" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6542-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="10" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6108-6112" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="9" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-6836" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  !<?xml version="1.0"?><Primers recycledIDs="5,4,3,0,2,1" nextValidID="19"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="6" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite></Primer><Primer recentID="7" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="9" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="10" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="11" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="12" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="13" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! 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  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="16:3:59">2019.5.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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(T{Nn+uPډe8wiiKOXg'J)RnS]zԩ8+kj=nNT7ڄdqNM*RJ[*e(foVT;AoP~fnF/ٻZ:WViEom9K%v
wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9kSJeMZeJgQf?^*U)CRfi]ՒVjjIIRieF"ZUYJ\Z]YxJje&&ҙyTVTTQ%bFIRIkiW iI)WeRMYeOe)djݖUc4sm`P'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ     l   s  "   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features nextValidID="22"><Feature recentID="0" name="mod rbs" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8302-8307" color="#a6acb3" type="standard"/></Feature><Feature recentID="21" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8974-8990" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="16" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="15" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="12" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="11" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9382-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="10" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8948-8952" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="9" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9676" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="8" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="7" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="6" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="5" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="4" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="3" name="FTL_1504" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8337-8947" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  ,<?xml version="1.0"?><Primers recycledIDs="2,1" nextValidID="19"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="KROL XXX FR2_R" sequence="atatGGTACCcttaacaccattgatagaagcgtttag" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:47:30Z"><BindingSite location="8920-8952" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8920-8952" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite><BindingSite simplified="1" location="8920-8952" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8920-8952" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite></Primer><Primer recentID="3" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="4" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="5" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="6" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="7" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="9" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="10" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="11" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="12" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="13" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="16" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="17" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="18" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  A<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="16:3:0">2019.5.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
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(T{Nn+uPډe8wiiKOXg'J)RnS]zԩ8+kj=nNT7ڄdqNM*RJ[*e(foVT;AoP~fnF/ٻZ:WViEom9K%v
wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9kSJeMZeJgQf?^*U)CRfi]ՒVjjIIRieF"ZUYJ\Z]YxJje&&ҙyTVTTQ%bFIRIkiW iI)WeRMYeOe)djݖUc4sm`P'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ     l   s  "   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features recycledIDs="0" nextValidID="22"><Feature recentID="21" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8974-8990" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="16" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="15" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="12" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="11" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9382-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="10" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8948-8952" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="9" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9676" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="8" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="7" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="6" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="5" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="4" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="3" name="FTL_1504" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8337-8947" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  2	<?xml version="1.0"?><Primers nextValidID="19"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="KROL XXX FR2_R" sequence="atatGGTACCcttaacaccattgatagaagcgtttag" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:47:30Z"><BindingSite location="8920-8952" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8920-8952" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite><BindingSite simplified="1" location="8920-8952" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8920-8952" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite></Primer><Primer recentID="1" name="KROL XXX FR2_F" sequence="atatTGCAGGAGGaaagcagttataattttctttagcatggt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:43:37Z"><BindingSite location="8307-8335" boundStrand="0" annealedBases="aaagcagttataattttctttagcatggt" meltingTemperature="57"><Component bases="atatTGCAGGAGG"/><Component hybridizedRange="8307-8335" bases="aaagcagttataattttctttagcatggt"/></BindingSite><BindingSite simplified="1" location="8307-8335" boundStrand="0" annealedBases="aaagcagttataattttctttagcatggt" meltingTemperature="57"><Component bases="atatTGCAGGAGG"/><Component hybridizedRange="8307-8335" bases="aaagcagttataattttctttagcatggt"/></BindingSite></Primer><Primer recentID="2" name="KROL XXX FR1_R" sequence="atatCCTCCTGCAgtctggaatgctactagcttctagc" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:41:03Z"><BindingSite location="8276-8300" boundStrand="1" annealedBases="gtctggaatgctactagcttctagc" meltingTemperature="58"><Component bases="atatCCTCCTGCA"/><Component hybridizedRange="8276-8300" bases="gtctggaatgctactagcttctagc"/></BindingSite><BindingSite simplified="1" location="8276-8300" boundStrand="1" annealedBases="gtctggaatgctactagcttctagc" meltingTemperature="58"><Component bases="atatCCTCCTGCA"/><Component hybridizedRange="8276-8300" bases="gtctggaatgctactagcttctagc"/></BindingSite></Primer><Primer recentID="3" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="4" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="5" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="6" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="7" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="9" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="10" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="11" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="12" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="13" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="16" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="17" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="18" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:42:42">2019.5.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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(T{Nn+uPډe8wiiKOXg'J)RnS]zԩ8+kj=nNT7ڄdqNM*RJ[*e(foVT;AoP~fnF/ٻZ:WViEom9K%v
wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9kASJeMZeJgQf?^*U)CRfi]ՒVjjIIRieF"ZUYJ\Z]YxJje&&ҙyTVTTQ%bFIRIkiW iI)WeRMYeOe)djݖUc4sm`P'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ     l   s  "   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features nextValidID="22"><Feature recentID="21" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8974-8990" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="16" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="15" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="12" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="11" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9382-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="10" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8948-8952" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="9" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9676" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="8" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="7" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="6" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="5" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="4" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="3" name="FTL_1504" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8337-8947" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="0" name="mod rbs" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8302-8307" color="#a6acb3" type="standard"/></Feature></Features>
  2d<?xml version="1.0"?><Primers nextValidID="19"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="KROL XXX FR2_R" sequence="atatGGTACCcttaacaccattgatagaagcgtttag" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:47:30Z"><BindingSite location="8920-8952" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8920-8952" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite><BindingSite simplified="1" location="8920-8952" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8920-8952" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite></Primer><Primer recentID="1" name="KROL XXX FR2_F" sequence="atatTGCAGGAGGaaagcagttataattttctttagcatggt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:43:37Z"><BindingSite location="8300-8335" boundStrand="0" annealedBases="CAGGAGGaaagcagttataattttctttagcatggt" meltingTemperature="62"><Component bases="atatTG"/><Component hybridizedRange="8300-8335" bases="CAGGAGGaaagcagttataattttctttagcatggt"/></BindingSite><BindingSite simplified="1" location="8300-8335" boundStrand="0" annealedBases="CAGGAGGaaagcagttataattttctttagcatggt" meltingTemperature="62"><Component bases="atatTG"/><Component hybridizedRange="8300-8335" bases="CAGGAGGaaagcagttataattttctttagcatggt"/></BindingSite></Primer><Primer recentID="2" name="KROL XXX FR1_R" sequence="atatCCTCCTGCAgtctggaatgctactagcttctagc" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:41:03Z"><BindingSite location="8276-8307" boundStrand="1" annealedBases="tCCTCCTGtctggaatgctactagcttctagc" meltingTemperature="59"><Component bases="ata"/><Component hybridizedRange="8300-8307" bases="tCCTCCTG"/><Component bases="CAg"/><Component hybridizedRange="8276-8299" bases="tctggaatgctactagcttctagc"/></BindingSite><BindingSite simplified="1" location="8276-8300" boundStrand="1" annealedBases="gtctggaatgctactagcttctagc" meltingTemperature="59"><Component bases="atatCCTCCTGCA"/><Component hybridizedRange="8276-8300" bases="gtctggaatgctactagcttctagc"/></BindingSite></Primer><Primer recentID="3" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="4" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="5" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="6" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="7" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="9" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="10" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="11" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="12" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="13" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="16" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="17" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="18" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:42:29">2019.5.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9kTҙj`RYfiשeJP锆WEjdFRj锤YiQVuE;jjiWVjYFIgIE^UUIXTFZkU(&@$+mmJUTSVEfYSyAceXM9\X"	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%     l   t  "   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features nextValidID="22"><Feature recentID="21" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8975-8991" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="16" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="15" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="12" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="11" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9383-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="10" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8949-8953" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="9" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9677" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="8" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="7" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="6" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="5" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="4" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="3" name="FTL_1504" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-8337" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8338-8948" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="0" name="mod rbs" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8303-8308" color="#a6acb3" type="standard"/></Feature></Features>
  2e<?xml version="1.0"?><Primers nextValidID="19"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="KROL XXX FR2_R" sequence="atatGGTACCcttaacaccattgatagaagcgtttag" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:47:30Z"><BindingSite location="8921-8953" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8921-8953" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite><BindingSite simplified="1" location="8921-8953" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8921-8953" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite></Primer><Primer recentID="1" name="KROL XXX FR2_F" sequence="atatTGCAGGAGGaaagcagttataattttctttagcatggt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:43:37Z"><BindingSite location="8301-8336" boundStrand="0" annealedBases="CAGGAGGaaagcagttataattttctttagcatggt" meltingTemperature="62"><Component bases="atatTG"/><Component hybridizedRange="8301-8336" bases="CAGGAGGaaagcagttataattttctttagcatggt"/></BindingSite><BindingSite simplified="1" location="8301-8336" boundStrand="0" annealedBases="CAGGAGGaaagcagttataattttctttagcatggt" meltingTemperature="62"><Component bases="atatTG"/><Component hybridizedRange="8301-8336" bases="CAGGAGGaaagcagttataattttctttagcatggt"/></BindingSite></Primer><Primer recentID="2" name="KROL XXX FR1_R" sequence="atatCCTCCTGCAgtctggaatgctactagcttctagc" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:41:03Z"><BindingSite location="8276-8308" boundStrand="1" annealedBases="tCCTCCTGgtctggaatgctactagcttctagc" meltingTemperature="60"><Component bases="ata"/><Component hybridizedRange="8301-8308" bases="tCCTCCTG"/><Component bases="CA"/><Component hybridizedRange="8276-8300" bases="gtctggaatgctactagcttctagc"/></BindingSite><BindingSite simplified="1" location="8276-8300" boundStrand="1" annealedBases="gtctggaatgctactagcttctagc" meltingTemperature="60"><Component bases="atatCCTCCTGCA"/><Component hybridizedRange="8276-8300" bases="gtctggaatgctactagcttctagc"/></BindingSite></Primer><Primer recentID="3" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="4" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="5" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="6" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="7" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="9" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="10" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="11" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="12" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="13" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="16" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="17" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9002-9021" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9002-9021" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="9002-9021" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9002-9021" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="18" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:41:34">2019.5.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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(T{Nn+uPډe8wiiKOXg'J)RnS]zԩ8+kj=nNT7ڄdqNM*RJ[*e(foVT;AoP~fnF/ٻZ:WViEom9K%v
wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9kAM)6	j9(JFn階VzVTHitVKVIY$e&JI 5jTSVYf+aqjufI)h5dVtJdUQZQQD%Jnda%^RdBQ$^KU5dVje=9vYUTӕι)@k%-"kgM"T>XHN^ݥ(?:,~,N63 D0ɠ3󯳷,8	PY\T]MT=5PU2?MUls-teF}N<gL:d$&+N|>     l   v  "   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features nextValidID="22"><Feature recentID="21" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8977-8993" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="16" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="15" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="12" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="11" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9385-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="10" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8951-8955" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="9" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9679" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="8" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="7" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="6" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="5" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="4" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="3" name="FTL_1504" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-8339" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8340-8950" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="0" name="mod rbs" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8305-8310" color="#a6acb3" type="standard"/></Feature></Features>
  2/<?xml version="1.0"?><Primers nextValidID="19"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="KROL XXX FR2_R" sequence="atatGGTACCcttaacaccattgatagaagcgtttag" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:47:30Z"><BindingSite location="8923-8955" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8923-8955" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite><BindingSite simplified="1" location="8923-8955" boundStrand="1" annealedBases="GGTACCcttaacaccattgatagaagcgtttag" meltingTemperature="62"><Component bases="atat"/><Component hybridizedRange="8923-8955" bases="GGTACCcttaacaccattgatagaagcgtttag"/></BindingSite></Primer><Primer recentID="1" name="KROL XXX FR2_F" sequence="atatTGCAGGAGGaaagcagttataattttctttagcatggt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:43:37Z"><BindingSite location="8301-8338" boundStrand="0" annealedBases="TGCAGGAGGaaagcagttataattttctttagcatggt" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="8301-8338" bases="TGCAGGAGGaaagcagttataattttctttagcatggt"/></BindingSite><BindingSite simplified="1" location="8301-8338" boundStrand="0" annealedBases="TGCAGGAGGaaagcagttataattttctttagcatggt" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="8301-8338" bases="TGCAGGAGGaaagcagttataattttctttagcatggt"/></BindingSite></Primer><Primer recentID="2" name="KROL XXX FR1_R" sequence="atatCCTCCTGCAgtctggaatgctactagcttctagc" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-21T20:41:03Z"><BindingSite location="8276-8310" boundStrand="1" annealedBases="tCCTCCTGCAgtctggaatgctactagcttctagc" meltingTemperature="67"><Component bases="ata"/><Component hybridizedRange="8276-8310" bases="tCCTCCTGCAgtctggaatgctactagcttctagc"/></BindingSite><BindingSite simplified="1" location="8276-8310" boundStrand="1" annealedBases="tCCTCCTGCAgtctggaatgctactagcttctagc" meltingTemperature="67"><Component bases="ata"/><Component hybridizedRange="8276-8310" bases="tCCTCCTGCAgtctggaatgctactagcttctagc"/></BindingSite></Primer><Primer recentID="3" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="4" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="5" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="6" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="7" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="9" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="10" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="11" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="12" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="13" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="16" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="17" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9004-9023" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9004-9023" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="9004-9023" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9004-9023" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="18" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:41:16">2019.5.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9k4Z%T*uj9ZCbYR:e!fZ-Y%fe)&ZT`(ժQNZedŪU՚$QiYm)WEjEEV$d)yJ		
[iDzfU-TՑYT^PbFjmeV9SNW:|NŬضN*Vo;uw]O6TSa!9{v;x=8X[\£&ξ3L&ӷC	g
ot[@sAQv!5PC_%T<82Cq6UԴBWy2;#;+ϫ2퓷8"B8"7o	     l   u  "   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
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consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="34" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="33" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="32" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="17" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="16" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="31" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9678" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="30" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8950-8954" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="29" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9384-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="28" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="27" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="26" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="25" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="24" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="23" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8976-8992" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  )<?xml version="1.0"?><Primers recycledIDs="17,16,10,7,1" nextValidID="21"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="11" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="12" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="13" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="2" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="3" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="4" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="5" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="6" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="9" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="18" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="19" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9003-9022" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9003-9022" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="9003-9022" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9003-9022" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="20" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="20:36:34">2019.5.21</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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(T{Nn+uPډe8wiiKOXg'J)RnS]zԩ8+kj=nNT7ڄdqNM*RJ[*e(foVT;AoP~fnF/ٻZ:WViEom9K%v
wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9kASJeMZeJgQf?^*U)CRfi]ՒVjjIIRieF"ZUYJ\Z]YxJje&&ҙyTVTTQ%bFIRIkiW iI)WeRMYeOe)djݖUc4sm`P'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ     l   s  "   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
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consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="34" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="33" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="32" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="17" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="16" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="31" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9676" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="30" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8948-8952" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="29" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9382-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="28" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="27" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="26" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="25" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="24" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="23" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8974-8990" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  )<?xml version="1.0"?><Primers recycledIDs="17,16,10,7,1" nextValidID="21"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="11" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="12" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="13" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="2" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="3" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="4" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="5" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="6" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="9" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="18" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="19" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="20" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="20:36:6">2019.5.21</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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(T{Nn+uPډe8wiiKOXg'J)RnS]zԩ8+kj=nNT7ڄdqNM*RJ[*e(foVT;AoP~fnF/ٻZ:WViEom9K%v
wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9k4Z%T*uj9ZCbYR:e!fZ-Y%fe)&ZT`(ժQNZedŪU՚$QiYm)WEjEEV$d)yJ		
[iDzfU-TՑYT^PbFjmeV9SNW:|NŬضN*Vo;uw]O6TSa!9{v;x=8X[\£&ξ3L&ӷC	g
ot[@sAQv!5PC_%T<82Cq6UԴBWy2;#;+ϫ2퓷8"B8"7o	    "   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  G<?xml version="1.0"?><Features recycledIDs="1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,18" nextValidID="37"><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8331-8941" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="36" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-8330" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="35" name="FTL_1504" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="34" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="33" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="32" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="17" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="16" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="31" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9670" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="30" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8942-8946" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="29" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9376-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="28" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="27" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="26" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="25" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="24" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="23" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8968-8984" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  )<?xml version="1.0"?><Primers recycledIDs="17,16,10,7,1" nextValidID="21"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="11" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="12" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="13" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="2" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="3" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="4" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="5" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="6" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="9" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="18" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="19" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="8995-9014" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="8995-9014" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="8995-9014" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="8995-9014" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="20" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="20:19:47">2019.5.21</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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(T{Nn+uPډe8wiiKOXg'J)RnS]zԩ8+kj=nNT7ڄdqNM*RJ[*e(foVT;AoP~fnF/ٻZ:WViEom9K%v
wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9kSJeMZeJgQf?^*U)CRfi]ՒVjjIIRieF"ZUYJ\Z]YxJje&&ҙyTVTTQ%bFIRIkiW iI)WeRMYeOe)djݖUc4sm`P'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ    "   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  G<?xml version="1.0"?><Features recycledIDs="1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,18" nextValidID="37"><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8337-8947" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="36" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="35" name="FTL_1504" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="34" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="33" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="32" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="17" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="16" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="31" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9676" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="30" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8948-8952" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="29" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9382-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="28" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="27" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="26" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="25" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="24" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="23" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8974-8990" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  )<?xml version="1.0"?><Primers recycledIDs="17,16,10,7,1" nextValidID="21"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="11" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="12" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="13" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="2" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="3" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="4" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="5" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="6" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="9" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="18" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="19" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="9001-9020" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="9001-9020" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="20" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="20:19:37">2019.5.21</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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(T{Nn+uPډe8wiiKOXg'J)RnS]zԩ8+kj=nNT7ڄdqNM*RJ[*e(foVT;AoP~fnF/ٻZ:WViEom9K%v
wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9kSJeM	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%        -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  8<?xml version="1.0"?><Features recycledIDs="0,1,2,3,4,5,6,7,8,9,10,11,12,13,14,15,18" nextValidID="37"><Feature recentID="36" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="35" name="FTL_1504" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="34" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="33" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="32" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="17" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="16" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="31" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9065" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="30" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8337-8341" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="29" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8771-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="28" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="27" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="26" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="25" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="24" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="23" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8363-8379" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  )<?xml version="1.0"?><Primers recycledIDs="17,16,10,7,1" nextValidID="21"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="11" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="12" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="13" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="2" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="3" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="4" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="5" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="6" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="8" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="9" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="18" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="19" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="8390-8409" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="8390-8409" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="8390-8409" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="8390-8409" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="20" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="20:18:45">2019.5.21</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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(T{Nn+uPډe8wiiKOXg'J)RnS]zԩ8+kj=nNT7ڄdqNM*RJ[*e(foVT;AoP~fnF/ٻZ:WViEom9K%v
wA}j$4R*J%BVkR$RQ&}=#~;Ymнhz}}X|֐1ҮPqjN7*}fi?ڈMJny!JߣˮM_CEtNUմ}t}h=J>ivӚ E/qϾfkcdeEAZ=VCKFٺSnJ:d[9:jVensm&d9kSJeMcYgmQ*}z&z}zod$#oZ}Mu~ikMq+e逸iڡj&6[J;RfUUVƩzUeٙe*jS@fUYyogY*6f~*	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%    #   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  2W<?xml version="1.0"?><Features nextValidID="37"><Feature recentID="36" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="35" name="FTL_1504" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-8294" color="#00e600" type="standard"/></Feature><Feature recentID="34" name="peroxidase/catalase" directionality="2" translationMW="81227.07" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6108-8294" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="33" name="P171 RT-FTL1504-1F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7507-7530" color="#0080ff" type="standard"/><Q name="note"><V text="P171 RT-FTL1504-1F"/></Q></Feature><Feature recentID="32" name="P172 RT-FTL1504-1R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7433-7456" color="#0080ff" type="standard"/><Q name="note"><V text="P172 RT-FTL1504-1R"/></Q></Feature><Feature recentID="17" name="P173 RT-FTL1504-2F" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6846-6869" color="#0080ff" type="standard"/><Q name="note"><V text="P173 RT-FTL1504-2F"/></Q></Feature><Feature recentID="16" name="P174 RT-FTL1504-2R" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6673-6696" color="#0080ff" type="standard"/><Q name="note"><V text="P174 RT-FTL1504-2R"/></Q></Feature><Feature recentID="31" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-9693" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="30" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="8965-8969" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="29" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="9399-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="28" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="27" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="26" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="25" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="24" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="23" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8991-9007" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8337-8819" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="15" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8339-8626" color="#00e600" type="standard"/></Feature><Feature recentID="14" name="co-chaperonin GroES" directionality="2" translationMW="10271.90" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="8339-8626" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8617-8644" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="12" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8589-8615" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="11" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8359-8385" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="10" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8682-8752" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="9" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8581-8819" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="8" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8746-8751" color="#a6acb3" type="standard"/></Feature><Feature recentID="7" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8725-8730" color="#a6acb3" type="standard"/></Feature><Feature recentID="6" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8718-8718" color="#ffffff" type="standard"/></Feature><Feature recentID="5" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8820-8964" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="4" name="FTL_1716" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8884-8964" color="#00e600" type="standard"/></Feature><Feature recentID="3" name="hypothetical protein" directionality="1" translationMW="3167.87" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="8997" maxFusedRunOn="8997"><Segment range="8884-8964" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="2" name="P052 groESpromoF" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8837-8865" color="#0080ff" type="standard"/><Q name="note"><V text="P052 groESpromoF"/></Q></Feature><Feature recentID="1" name="P262 groESpCheck2" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8896-8921" color="#8080ff" type="standard"/><Q name="note"><V text="P262 groESpCheck2"/></Q></Feature><Feature recentID="0" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="8820-8864" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature></Features>
  HP<?xml version="1.0"?><Primers nextValidID="21"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P174 RT-FTL1504-2R" sequence="GCTGCTTGCTCAACACCCACATTA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite><BindingSite simplified="1" location="6672-6695" boundStrand="0" annealedBases="GCTGCTTGCTCAACACCCACATTA" meltingTemperature="59"><Component hybridizedRange="6672-6695" bases="GCTGCTTGCTCAACACCCACATTA"/></BindingSite></Primer><Primer recentID="11" name="P173 RT-FTL1504-2F" sequence="GGTGGCTCAAATGGTGCAAGGATT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite><BindingSite simplified="1" location="6845-6868" boundStrand="1" annealedBases="GGTGGCTCAAATGGTGCAAGGATT" meltingTemperature="62"><Component hybridizedRange="6845-6868" bases="GGTGGCTCAAATGGTGCAAGGATT"/></BindingSite></Primer><Primer recentID="12" name="P172 RT-FTL1504-1R" sequence="TATTGTGCCAGCCTAGACCTTGCT" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite><BindingSite simplified="1" location="7432-7455" boundStrand="0" annealedBases="TATTGTGCCAGCCTAGACCTTGCT" meltingTemperature="62"><Component hybridizedRange="7432-7455" bases="TATTGTGCCAGCCTAGACCTTGCT"/></BindingSite></Primer><Primer recentID="13" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="7506-7529" boundStrand="1" annealedBases="AACTCATGGTGCAGTTCCAGAGGA" meltingTemperature="62"><Component hybridizedRange="7506-7529" bases="AACTCATGGTGCAGTTCCAGAGGA"/></BindingSite></Primer><Primer recentID="1" name="KROL132 GroEL rbs FR2 R" sequence="atatGGTACCaaacatcgcaaaaggtgtaaatattataa" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="8935-8969" boundStrand="1" annealedBases="GGTACCaaacatcgcaaaaggtgtaaatattataa" meltingTemperature="59"><Component bases="atat"/><Component hybridizedRange="8935-8969" bases="GGTACCaaacatcgcaaaaggtgtaaatattataa"/></BindingSite><BindingSite simplified="1" location="8935-8969" boundStrand="1" annealedBases="GGTACCaaacatcgcaaaaggtgtaaatattataa" meltingTemperature="59"><Component bases="atat"/><Component hybridizedRange="8935-8969" bases="GGTACCaaacatcgcaaaaggtgtaaatattataa"/></BindingSite></Primer><Primer recentID="2" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="8836-8863" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="8836-8863" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="8836-8863" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="8836-8863" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="3" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="8836-8864" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCC"/><Component hybridizedRange="8836-8864" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="8836-8864" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCC"/><Component hybridizedRange="8836-8864" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="4" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="8836-8864" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="8836-8864" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="8836-8864" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="8836-8864" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="5" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="8836-8863" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="8836-8863" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="8836-8863" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="8836-8863" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="6" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="8836-8863" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCATATG"/><Component hybridizedRange="8836-8863" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="8836-8863" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCATATG"/><Component hybridizedRange="8836-8863" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="7" name="P736 IntCompLon1F" sequence="atatCTGCAGttgtatggattagtcgagctaaaaagc" description="Contains PstI site"><BindingSite location="8887-8913" boundStrand="1" annealedBases="ttgtatggattagtcgagctaaaaagc" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="8887-8913" bases="ttgtatggattagtcgagctaaaaagc"/></BindingSite><BindingSite simplified="1" location="8887-8913" boundStrand="1" annealedBases="ttgtatggattagtcgagctaaaaagc" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="8887-8913" bases="ttgtatggattagtcgagctaaaaagc"/></BindingSite></Primer><Primer recentID="8" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="8831-8858" boundStrand="0" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="8831-8858" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="8831-8858" boundStrand="0" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="8831-8858" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="9" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="8832-8859" boundStrand="0" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="8832-8859" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="8832-8859" boundStrand="0" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="8832-8859" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="10" name="P262 groPChk2" sequence="aatcctattgtatggattagtcgagc" description="0"><BindingSite location="8895-8920" boundStrand="1" annealedBases="aatcctattgtatggattagtcgagc" meltingTemperature="57"><Component hybridizedRange="8895-8920" bases="aatcctattgtatggattagtcgagc"/></BindingSite><BindingSite simplified="1" location="8895-8920" boundStrand="1" annealedBases="aatcctattgtatggattagtcgagc" meltingTemperature="57"><Component hybridizedRange="8895-8920" bases="aatcctattgtatggattagtcgagc"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="8610-8643" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="8610-8614" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="8615-8643" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="8615-8643" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="8615-8643" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! 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  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="20:17:56">2019.5.21</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="24" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="23" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6762-6778" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6590" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="15" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6110-6397" color="#00e600" type="standard"/></Feature><Feature recentID="14" name="co-chaperonin GroES" directionality="2" translationMW="10271.90" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6110-6397" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6388-6415" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="12" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6360-6386" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="11" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6130-6156" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="10" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6453-6523" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="9" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6352-6590" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="8" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6517-6522" color="#a6acb3" type="standard"/></Feature><Feature recentID="7" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6496-6501" color="#a6acb3" type="standard"/></Feature><Feature recentID="6" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6489-6489" color="#ffffff" type="standard"/></Feature><Feature recentID="5" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6591-6735" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="4" name="FTL_1716" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6655-6735" color="#00e600" type="standard"/></Feature><Feature recentID="3" name="hypothetical protein" directionality="1" translationMW="3167.87" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="6768" maxFusedRunOn="6768"><Segment range="6655-6735" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="2" name="P052 groESpromoF" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6608-6636" color="#0080ff" type="standard"/><Q name="note"><V text="P052 groESpromoF"/></Q></Feature><Feature recentID="1" name="P262 groESpCheck2" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6667-6692" color="#8080ff" type="standard"/><Q name="note"><V text="P262 groESpCheck2"/></Q></Feature><Feature recentID="0" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6591-6635" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature></Features>
  =g<?xml version="1.0"?><Primers recycledIDs="13,12,11,0" nextValidID="21"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="1" name="KROL132 GroEL rbs FR2 R" sequence="atatGGTACCaaacatcgcaaaaggtgtaaatattataa" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6706-6740" boundStrand="1" annealedBases="GGTACCaaacatcgcaaaaggtgtaaatattataa" meltingTemperature="59"><Component bases="atat"/><Component hybridizedRange="6706-6740" bases="GGTACCaaacatcgcaaaaggtgtaaatattataa"/></BindingSite><BindingSite simplified="1" location="6706-6740" boundStrand="1" annealedBases="GGTACCaaacatcgcaaaaggtgtaaatattataa" meltingTemperature="59"><Component bases="atat"/><Component hybridizedRange="6706-6740" bases="GGTACCaaacatcgcaaaaggtgtaaatattataa"/></BindingSite></Primer><Primer recentID="2" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="6607-6634" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="6607-6634" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="6607-6634" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="6607-6634" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="3" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="6607-6635" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCC"/><Component hybridizedRange="6607-6635" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="6607-6635" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCC"/><Component hybridizedRange="6607-6635" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="4" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="6607-6635" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="6607-6635" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="6607-6635" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="6607-6635" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="5" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="6607-6634" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="6607-6634" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="6607-6634" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="6607-6634" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="6" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="6607-6634" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCATATG"/><Component hybridizedRange="6607-6634" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="6607-6634" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCATATG"/><Component hybridizedRange="6607-6634" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="7" name="P736 IntCompLon1F" sequence="atatCTGCAGttgtatggattagtcgagctaaaaagc" description="Contains PstI site"><BindingSite location="6658-6684" boundStrand="1" annealedBases="ttgtatggattagtcgagctaaaaagc" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="6658-6684" bases="ttgtatggattagtcgagctaaaaagc"/></BindingSite><BindingSite simplified="1" location="6658-6684" boundStrand="1" annealedBases="ttgtatggattagtcgagctaaaaagc" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="6658-6684" bases="ttgtatggattagtcgagctaaaaagc"/></BindingSite></Primer><Primer recentID="8" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="6602-6629" boundStrand="0" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="6602-6629" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="6602-6629" boundStrand="0" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="6602-6629" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="9" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="6603-6630" boundStrand="0" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="6603-6630" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="6603-6630" boundStrand="0" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="6603-6630" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="10" name="P262 groPChk2" sequence="aatcctattgtatggattagtcgagc" description="0"><BindingSite location="6666-6691" boundStrand="1" annealedBases="aatcctattgtatggattagtcgagc" meltingTemperature="57"><Component hybridizedRange="6666-6691" bases="aatcctattgtatggattagtcgagc"/></BindingSite><BindingSite simplified="1" location="6666-6691" boundStrand="1" annealedBases="aatcctattgtatggattagtcgagc" meltingTemperature="57"><Component hybridizedRange="6666-6691" bases="aatcctattgtatggattagtcgagc"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="6381-6414" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="6381-6385" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="6386-6414" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6386-6414" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="6386-6414" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="6387-6414" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6387-6414" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6387-6414" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6387-6414" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="16" name="P224 SP_p1715_F" sequence="tatcttcttctctcatcattagaagag" description="0"><BindingSite location="6129-6155" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="54"><Component hybridizedRange="6129-6155" bases="tatcttcttctctcatcattagaagag"/></BindingSite><BindingSite simplified="1" location="6129-6155" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="54"><Component hybridizedRange="6129-6155" bases="tatcttcttctctcatcattagaagag"/></BindingSite></Primer><Primer recentID="17" name="P225 SP_p1715_R" sequence="tccattacaagatagagtattagttcg" description="0"><BindingSite location="6359-6385" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="55"><Component hybridizedRange="6359-6385" bases="tccattacaagatagagtattagttcg"/></BindingSite><BindingSite simplified="1" location="6359-6385" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="55"><Component hybridizedRange="6359-6385" bases="tccattacaagatagagtattagttcg"/></BindingSite></Primer><Primer recentID="18" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="6538-6564" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="6538-6564" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="6538-6564" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="6538-6564" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="19" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6789-6808" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="6789-6808" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="6789-6808" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="6789-6808" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="20" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="20:17:26">2019.5.21</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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6JQѪ7^Yn`qb]nVMcnuTS5.FhKUUZEj*-]NjiU^NV]ZR:	*eUNNǖujijwbk%-"kgM"T>XHN^ݥ(?:,~,N63 D0ɠ3󯳷,8	PY\T]MT=5PU2?MUls-teF}N<gL:d$&+N|>        e   -<AlignableSequences trimStringency="Medium"/>  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  /.<?xml version="1.0"?><Features nextValidID="32"><Feature recentID="31" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7999" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="30" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7271-7275" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="29" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7705-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="28" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="27" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="26" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="25" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="24" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="23" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7297-7313" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="18" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7125" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="17" name="groEL" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6607" color="#00e600" type="standard"/></Feature><Feature recentID="16" name="chaperonin GroEL" directionality="2" translationMW="17231.43" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="5858" maxFusedRunOn="5858"><Segment range="6108-6607" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="60 kDa chaperone family; promotes refolding of misfolded polypeptides especially under stressful conditions; forms two stacked rings of heptamers to form a barrel-shaped 14mer; ends can be capped by GroES; misfolded proteins enter the barrel where they are refolded when GroES binds; many bacteria have multiple copies of the groEL gene which are active under different environmental conditions; the B.japonicum protein in this cluster is expressed constitutively; in Rhodobacter, Corynebacterium and Rhizobium this protein is essential for growth"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="15" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6645-6932" color="#00e600" type="standard"/></Feature><Feature recentID="14" name="co-chaperonin GroES" directionality="2" translationMW="10272.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6645-6932" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="13" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6923-6950" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="12" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6895-6921" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="11" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6665-6691" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="10" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6988-7058" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="9" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6887-7125" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="8" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7052-7057" color="#a6acb3" type="standard"/></Feature><Feature recentID="7" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7031-7036" color="#a6acb3" type="standard"/></Feature><Feature recentID="6" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7024-7024" color="#ffffff" type="standard"/></Feature><Feature recentID="5" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7126-7270" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="4" name="FTL_1716" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7190-7270" color="#00e600" type="standard"/></Feature><Feature recentID="3" name="hypothetical protein" directionality="1" translationMW="3167.93" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="7303" maxFusedRunOn="7303"><Segment range="7190-7270" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="2" name="P052 groESpromoF" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7143-7171" color="#0080ff" type="standard"/><Q name="note"><V text="P052 groESpromoF"/></Q></Feature><Feature recentID="1" name="P262 groESpCheck2" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7202-7227" color="#8080ff" type="standard"/><Q name="note"><V text="P262 groESpCheck2"/></Q></Feature><Feature recentID="0" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7126-7170" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature></Features>
  G<?xml version="1.0"?><Primers nextValidID="21"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="KROL133 GroEL rbs seq primer" sequence="cctaaagtaacttttacagcatttgct" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6512-6538" boundStrand="0" annealedBases="cctaaagtaacttttacagcatttgct" meltingTemperature="56"><Component hybridizedRange="6512-6538" bases="cctaaagtaacttttacagcatttgct"/></BindingSite><BindingSite simplified="1" location="6512-6538" boundStrand="0" annealedBases="cctaaagtaacttttacagcatttgct" meltingTemperature="56"><Component hybridizedRange="6512-6538" bases="cctaaagtaacttttacagcatttgct"/></BindingSite></Primer><Primer recentID="1" name="KROL132 GroEL rbs FR2 R" sequence="atatGGTACCaaacatcgcaaaaggtgtaaatattataa" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7241-7275" boundStrand="1" annealedBases="GGTACCaaacatcgcaaaaggtgtaaatattataa" meltingTemperature="59"><Component bases="atat"/><Component hybridizedRange="7241-7275" bases="GGTACCaaacatcgcaaaaggtgtaaatattataa"/></BindingSite><BindingSite simplified="1" location="7241-7275" boundStrand="1" annealedBases="GGTACCaaacatcgcaaaaggtgtaaatattataa" meltingTemperature="59"><Component bases="atat"/><Component hybridizedRange="7241-7275" bases="GGTACCaaacatcgcaaaaggtgtaaatattataa"/></BindingSite></Primer><Primer recentID="2" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="52"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="7142-7169" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="52"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="7142-7169" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="52"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="7142-7169" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="52"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="7142-7169" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="3" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="7142-7170" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCC"/><Component hybridizedRange="7142-7170" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="7142-7170" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCC"/><Component hybridizedRange="7142-7170" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="4" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="7142-7170" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="7142-7170" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="7142-7170" boundStrand="1" annealedBases="atatacccttcaagctttgaaaaataaac" meltingTemperature="56"><Component bases="atatCTGCAG"/><Component hybridizedRange="7142-7170" bases="atatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="5" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="7142-7169" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="52"><Component bases="atatCTCGAG"/><Component hybridizedRange="7142-7169" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="7142-7169" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="52"><Component bases="atatCTCGAG"/><Component hybridizedRange="7142-7169" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="6" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite location="7142-7169" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="52"><Component bases="atatCATATG"/><Component hybridizedRange="7142-7169" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="7142-7169" boundStrand="1" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="52"><Component bases="atatCATATG"/><Component hybridizedRange="7142-7169" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="7" name="P736 IntCompLon1F" sequence="atatCTGCAGttgtatggattagtcgagctaaaaagc" description="Contains PstI site"><BindingSite location="7193-7219" boundStrand="1" annealedBases="ttgtatggattagtcgagctaaaaagc" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="7193-7219" bases="ttgtatggattagtcgagctaaaaagc"/></BindingSite><BindingSite simplified="1" location="7193-7219" boundStrand="1" annealedBases="ttgtatggattagtcgagctaaaaagc" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="7193-7219" bases="ttgtatggattagtcgagctaaaaagc"/></BindingSite></Primer><Primer recentID="8" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="7137-7164" boundStrand="0" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="7137-7164" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="7137-7164" boundStrand="0" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="7137-7164" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="9" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="7138-7165" boundStrand="0" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="55"><Component hybridizedRange="7138-7165" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="55"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="7138-7165" boundStrand="0" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="55"><Component hybridizedRange="7138-7165" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="55"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="10" name="P262 groPChk2" sequence="aatcctattgtatggattagtcgagc" description="0"><BindingSite location="7201-7226" boundStrand="1" annealedBases="aatcctattgtatggattagtcgagc" meltingTemperature="57"><Component hybridizedRange="7201-7226" bases="aatcctattgtatggattagtcgagc"/></BindingSite><BindingSite simplified="1" location="7201-7226" boundStrand="1" annealedBases="aatcctattgtatggattagtcgagc" meltingTemperature="57"><Component hybridizedRange="7201-7226" bases="aatcctattgtatggattagtcgagc"/></BindingSite></Primer><Primer recentID="11" name="KROL131 GroEL FR2 F" sequence="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6605-6650" boundStrand="0" annealedBases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" meltingTemperature="60"><Component hybridizedRange="6605-6650" bases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca"/></BindingSite><BindingSite simplified="1" location="6605-6650" boundStrand="0" annealedBases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" meltingTemperature="60"><Component hybridizedRange="6605-6650" bases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca"/></BindingSite></Primer><Primer recentID="12" name="KROL130 GroEL rbs FR1 F" sequence="atatGGATCCgccattgcgtcagcgataagctta" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6101-6130" boundStrand="0" annealedBases="GGATCCgccattgcgtcagcgataagctta" meltingTemperature="65"><Component bases="atat"/><Component hybridizedRange="6101-6130" bases="GGATCCgccattgcgtcagcgataagctta"/></BindingSite><BindingSite simplified="1" location="6101-6130" boundStrand="0" annealedBases="GGATCCgccattgcgtcagcgataagctta" meltingTemperature="65"><Component bases="atat"/><Component hybridizedRange="6101-6130" bases="GGATCCgccattgcgtcagcgataagctta"/></BindingSite></Primer><Primer recentID="13" name="KROL129 GroEL rbs FR1 R" sequence="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6586-6630" boundStrand="1" annealedBases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" meltingTemperature="62"><Component hybridizedRange="6586-6630" bases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta"/></BindingSite><BindingSite simplified="1" location="6586-6630" boundStrand="1" annealedBases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" meltingTemperature="62"><Component hybridizedRange="6586-6630" bases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta"/></BindingSite></Primer><Primer recentID="14" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="6916-6949" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="6916-6920" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6921-6949" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="15" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="16" name="P224 SP_p1715_F" sequence="tatcttcttctctcatcattagaagag" description="0"><BindingSite location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite><BindingSite simplified="1" location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite></Primer><Primer recentID="17" name="P225 SP_p1715_R" sequence="tccattacaagatagagtattagttcg" description="0"><BindingSite location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite><BindingSite simplified="1" location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite></Primer><Primer recentID="18" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="19" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7324-7343" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7324-7343" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7324-7343" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7324-7343" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="20" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="20:17:17">2019.5.21</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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6JQѪ7^Yn`qb]nVMcnuTS5.FhKUUZEj*-]NjiU^NVB|NŬضN*Vo;uw]O6TSa!9{v;x=8X[\£&ξ3L&ӷC	g
ot[@sAQv!5PC_%T<82Cq6UԴBWy2;#;+ϫ2퓷8"B8"7o	          !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  (M<?xml version="1.0"?><Features nextValidID="26"><Feature recentID="25" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7024-7024" color="#ffffff" type="standard"/></Feature><Feature recentID="24" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7031-7036" color="#a6acb3" type="standard"/></Feature><Feature recentID="23" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7052-7057" color="#a6acb3" type="standard"/></Feature><Feature recentID="22" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6887-7125" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="21" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6988-7058" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="20" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6665-6691" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="19" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6895-6921" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="18" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6923-6950" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="17" name="co-chaperonin GroES" directionality="2" translationMW="10272.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6645-6932" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="3" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6645-6932" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="chaperonin GroEL" directionality="2" translationMW="17231.43" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="5858" maxFusedRunOn="5858"><Segment range="6108-6607" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="60 kDa chaperone family; promotes refolding of misfolded polypeptides especially under stressful conditions; forms two stacked rings of heptamers to form a barrel-shaped 14mer; ends can be capped by GroES; misfolded proteins enter the barrel where they are refolded when GroES binds; many bacteria have multiple copies of the groEL gene which are active under different environmental conditions; the B.japonicum protein in this cluster is expressed constitutively; in Rhodobacter, Corynebacterium and Rhizobium this protein is essential for growth"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="groEL" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6607" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7125" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="16" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7152-7168" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="10" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="6" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7560-288" color="#ffff00" type="standard"/><Q name="note"><V text="ColE origin"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="5" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7126-7130" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="4" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7854" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  ?<?xml version="1.0"?><Primers recycledIDs="10" nextValidID="11"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="9" name="KROL131 GroES FR2 F" sequence="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6605-6650" boundStrand="0" annealedBases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" meltingTemperature="60"><Component hybridizedRange="6605-6650" bases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca"/></BindingSite><BindingSite simplified="1" location="6605-6650" boundStrand="0" annealedBases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" meltingTemperature="60"><Component hybridizedRange="6605-6650" bases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca"/></BindingSite></Primer><Primer recentID="8" name="KROL130 GroES FR1 F" sequence="gccattgcgtcagcgataagctta" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6107-6130" boundStrand="0" annealedBases="gccattgcgtcagcgataagctta" meltingTemperature="59"><Component hybridizedRange="6107-6130" bases="gccattgcgtcagcgataagctta"/></BindingSite><BindingSite simplified="1" location="6107-6130" boundStrand="0" annealedBases="gccattgcgtcagcgataagctta" meltingTemperature="59"><Component hybridizedRange="6107-6130" bases="gccattgcgtcagcgataagctta"/></BindingSite></Primer><Primer recentID="7" name="KROL129 GroES FR1 R" sequence="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6586-6630" boundStrand="1" annealedBases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" meltingTemperature="62"><Component hybridizedRange="6586-6630" bases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta"/></BindingSite><BindingSite simplified="1" location="6586-6630" boundStrand="1" annealedBases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" meltingTemperature="62"><Component hybridizedRange="6586-6630" bases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta"/></BindingSite></Primer><Primer recentID="0" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="6916-6949" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="6916-6920" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6921-6949" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="1" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="2" name="P224 SP_p1715_F" sequence="tatcttcttctctcatcattagaagag" description="0"><BindingSite location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite><BindingSite simplified="1" location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite></Primer><Primer recentID="3" name="P225 SP_p1715_R" sequence="tccattacaagatagagtattagttcg" description="0"><BindingSite location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite><BindingSite simplified="1" location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite></Primer><Primer recentID="4" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="5" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7179-7198" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7179-7198" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7179-7198" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7179-7198" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="6" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="15:38:24">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  U               .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
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ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NASI6)4FIhj]Tky~M(,gFA!ӪGYc5'K8IMvSٮjMtg}(3=XjnUiR^j9")wۓjq:ۅYWکhiVQ5aEתz}j}]7תvBL=}t_9
6JQѪ7^Yn`qb]nVMcnuTS5.FhKUUZEj*-]NjiU^NV\'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ          !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  (M<?xml version="1.0"?><Features nextValidID="26"><Feature recentID="25" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7024-7024" color="#ffffff" type="standard"/></Feature><Feature recentID="24" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7031-7036" color="#a6acb3" type="standard"/></Feature><Feature recentID="23" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7052-7057" color="#a6acb3" type="standard"/></Feature><Feature recentID="22" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6887-7127" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="21" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6988-7058" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="20" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6665-6691" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="19" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6895-6921" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="18" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6923-6950" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="17" name="co-chaperonin GroES" directionality="2" translationMW="10272.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6645-6932" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="3" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6645-6932" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="chaperonin GroEL" directionality="2" translationMW="17231.43" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="5858" maxFusedRunOn="5858"><Segment range="6108-6607" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="60 kDa chaperone family; promotes refolding of misfolded polypeptides especially under stressful conditions; forms two stacked rings of heptamers to form a barrel-shaped 14mer; ends can be capped by GroES; misfolded proteins enter the barrel where they are refolded when GroES binds; many bacteria have multiple copies of the groEL gene which are active under different environmental conditions; the B.japonicum protein in this cluster is expressed constitutively; in Rhodobacter, Corynebacterium and Rhizobium this protein is essential for growth"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="groEL" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6607" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7127" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="16" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7154-7170" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="10" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="6" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7562-288" color="#ffff00" type="standard"/><Q name="note"><V text="ColE origin"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="5" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7128-7132" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="4" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7856" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  "|<?xml version="1.0"?><Primers nextValidID="11"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="10" name="KROL132 GroES FR2 R" sequence="tgttatttagctagtttttttaattaaagtt" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="907-937" boundStrand="0" annealedBases="tgttatttagctagtttttttaattaaagtt" meltingTemperature="51"><Component hybridizedRange="907-937" bases="tgttatttagctagtttttttaattaaagtt"/></BindingSite><BindingSite location="7097-7126" boundStrand="1" annealedBases="gttatttagctagtttttttaattaaagtt" meltingTemperature="50"><Component bases="t"/><Component hybridizedRange="7097-7126" bases="gttatttagctagtttttttaattaaagtt"/></BindingSite><BindingSite simplified="1" location="907-937" boundStrand="0" annealedBases="tgttatttagctagtttttttaattaaagtt" meltingTemperature="51"><Component hybridizedRange="907-937" bases="tgttatttagctagtttttttaattaaagtt"/></BindingSite><BindingSite simplified="1" location="7097-7126" boundStrand="1" annealedBases="gttatttagctagtttttttaattaaagtt" meltingTemperature="50"><Component bases="t"/><Component hybridizedRange="7097-7126" bases="gttatttagctagtttttttaattaaagtt"/></BindingSite></Primer><Primer recentID="9" name="KROL131 GroES FR2 F" sequence="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6605-6650" boundStrand="0" annealedBases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" meltingTemperature="60"><Component hybridizedRange="6605-6650" bases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca"/></BindingSite><BindingSite simplified="1" location="6605-6650" boundStrand="0" annealedBases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" meltingTemperature="60"><Component hybridizedRange="6605-6650" bases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca"/></BindingSite></Primer><Primer recentID="8" name="KROL130 GroES FR1 F" sequence="gccattgcgtcagcgataagctta" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6107-6130" boundStrand="0" annealedBases="gccattgcgtcagcgataagctta" meltingTemperature="59"><Component hybridizedRange="6107-6130" bases="gccattgcgtcagcgataagctta"/></BindingSite><BindingSite simplified="1" location="6107-6130" boundStrand="0" annealedBases="gccattgcgtcagcgataagctta" meltingTemperature="59"><Component hybridizedRange="6107-6130" bases="gccattgcgtcagcgataagctta"/></BindingSite></Primer><Primer recentID="7" name="KROL129 GroES FR1 R" sequence="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6586-6630" boundStrand="1" annealedBases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" meltingTemperature="62"><Component hybridizedRange="6586-6630" bases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta"/></BindingSite><BindingSite simplified="1" location="6586-6630" boundStrand="1" annealedBases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" meltingTemperature="62"><Component hybridizedRange="6586-6630" bases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta"/></BindingSite></Primer><Primer recentID="0" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="6916-6949" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="6916-6920" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6921-6949" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="1" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="2" name="P224 SP_p1715_F" sequence="tatcttcttctctcatcattagaagag" description="0"><BindingSite location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite><BindingSite simplified="1" location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite></Primer><Primer recentID="3" name="P225 SP_p1715_R" sequence="tccattacaagatagagtattagttcg" description="0"><BindingSite location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite><BindingSite simplified="1" location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite></Primer><Primer recentID="4" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="5" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7181-7200" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7181-7200" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7181-7200" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7181-7200" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="6" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  A<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="15:35:8">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  T               .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
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ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NASI6)4FIhj]Tky~M(,gFA!ӪGYc5'K8IMvSٮjMtg}(3=XjnUiR^j9")wۓjq:ۅYWکhiVQ5aEתz}j}]7תvBL=}t_9
6JQѪ7^Yn`qb]nVMcnuTS5.FhKUUZEj*-]NjiU^NV]	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%          !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  (M<?xml version="1.0"?><Features nextValidID="26"><Feature recentID="25" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7024-7024" color="#ffffff" type="standard"/></Feature><Feature recentID="24" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7031-7036" color="#a6acb3" type="standard"/></Feature><Feature recentID="23" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7052-7057" color="#a6acb3" type="standard"/></Feature><Feature recentID="22" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6887-7128" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="21" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6988-7058" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="20" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6665-6691" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="19" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6895-6921" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="18" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6923-6950" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="17" name="co-chaperonin GroES" directionality="2" translationMW="10272.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6645-6932" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="3" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6645-6932" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="chaperonin GroEL" directionality="2" translationMW="17231.43" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="5858" maxFusedRunOn="5858"><Segment range="6108-6607" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="60 kDa chaperone family; promotes refolding of misfolded polypeptides especially under stressful conditions; forms two stacked rings of heptamers to form a barrel-shaped 14mer; ends can be capped by GroES; misfolded proteins enter the barrel where they are refolded when GroES binds; many bacteria have multiple copies of the groEL gene which are active under different environmental conditions; the B.japonicum protein in this cluster is expressed constitutively; in Rhodobacter, Corynebacterium and Rhizobium this protein is essential for growth"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="groEL" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6607" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7128" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="16" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7155-7171" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="10" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="6" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7563-288" color="#ffff00" type="standard"/><Q name="note"><V text="ColE origin"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="5" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7129-7133" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="4" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7857" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  "J<?xml version="1.0"?><Primers nextValidID="11"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="10" name="KROL132 GroES FR2 R" sequence="tgttatttagctagtttttttaattaaagtt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="907-937" boundStrand="0" annealedBases="tgttatttagctagtttttttaattaaagtt" meltingTemperature="51"><Component hybridizedRange="907-937" bases="tgttatttagctagtttttttaattaaagtt"/></BindingSite><BindingSite location="7097-7127" boundStrand="1" annealedBases="tgttatttagctagtttttttaattaaagtt" meltingTemperature="51"><Component hybridizedRange="7097-7127" bases="tgttatttagctagtttttttaattaaagtt"/></BindingSite><BindingSite simplified="1" location="907-937" boundStrand="0" annealedBases="tgttatttagctagtttttttaattaaagtt" meltingTemperature="51"><Component hybridizedRange="907-937" bases="tgttatttagctagtttttttaattaaagtt"/></BindingSite><BindingSite simplified="1" location="7097-7127" boundStrand="1" annealedBases="tgttatttagctagtttttttaattaaagtt" meltingTemperature="51"><Component hybridizedRange="7097-7127" bases="tgttatttagctagtttttttaattaaagtt"/></BindingSite></Primer><Primer recentID="9" name="KROL131 GroES FR2 F" sequence="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6605-6650" boundStrand="0" annealedBases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" meltingTemperature="60"><Component hybridizedRange="6605-6650" bases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca"/></BindingSite><BindingSite simplified="1" location="6605-6650" boundStrand="0" annealedBases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca" meltingTemperature="60"><Component hybridizedRange="6605-6650" bases="atttttaattGGAGGAttaaaatttgtaagattctttgattatgca"/></BindingSite></Primer><Primer recentID="8" name="KROL130 GroES FR1 F" sequence="gccattgcgtcagcgataagctta" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6107-6130" boundStrand="0" annealedBases="gccattgcgtcagcgataagctta" meltingTemperature="59"><Component hybridizedRange="6107-6130" bases="gccattgcgtcagcgataagctta"/></BindingSite><BindingSite simplified="1" location="6107-6130" boundStrand="0" annealedBases="gccattgcgtcagcgataagctta" meltingTemperature="59"><Component hybridizedRange="6107-6130" bases="gccattgcgtcagcgataagctta"/></BindingSite></Primer><Primer recentID="7" name="KROL129 GroES FR1 R" sequence="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6586-6630" boundStrand="1" annealedBases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" meltingTemperature="62"><Component hybridizedRange="6586-6630" bases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta"/></BindingSite><BindingSite simplified="1" location="6586-6630" boundStrand="1" annealedBases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta" meltingTemperature="62"><Component hybridizedRange="6586-6630" bases="caaattttaaTCCTCCaattaaaaatggctgcaaaacaagtttta"/></BindingSite></Primer><Primer recentID="0" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="6916-6949" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="6916-6920" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6921-6949" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="1" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="2" name="P224 SP_p1715_F" sequence="tatcttcttctctcatcattagaagag" description="0"><BindingSite location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite><BindingSite simplified="1" location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite></Primer><Primer recentID="3" name="P225 SP_p1715_R" sequence="tccattacaagatagagtattagttcg" description="0"><BindingSite location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite><BindingSite simplified="1" location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite></Primer><Primer recentID="4" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="5" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7182-7201" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7182-7201" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7182-7201" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7182-7201" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="6" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="15:32:57">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NASI6)4FIhj]Tky~M(,gFA!ӪGYc5'K8IMvSٮjMtg}(3=XjnUiR^j9")wۓjq:ۅYWکjj%cYgmQ*}z&z}zod$#oZ}Mu~ikMq+e逸iڡj&6[J;RfUUVƩzUeПk%-"kgM"T>XHN^ݥ(?:,~,N63 D0ɠ3󯳷,8	PY\T]MT=5PU2?MUls-teF}N<gL:d$&+N|>      !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  (M<?xml version="1.0"?><Features nextValidID="26"><Feature recentID="25" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7018-7018" color="#ffffff" type="standard"/></Feature><Feature recentID="24" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7025-7030" color="#a6acb3" type="standard"/></Feature><Feature recentID="23" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7046-7051" color="#a6acb3" type="standard"/></Feature><Feature recentID="22" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6881-7122" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="21" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6982-7052" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="20" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6659-6685" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="19" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6889-6915" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="18" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6917-6944" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="17" name="co-chaperonin GroES" directionality="2" translationMW="10272.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6639-6926" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="3" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6639-6926" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="chaperonin GroEL" directionality="2" translationMW="17231.43" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="5858" maxFusedRunOn="5858"><Segment range="6108-6607" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="60 kDa chaperone family; promotes refolding of misfolded polypeptides especially under stressful conditions; forms two stacked rings of heptamers to form a barrel-shaped 14mer; ends can be capped by GroES; misfolded proteins enter the barrel where they are refolded when GroES binds; many bacteria have multiple copies of the groEL gene which are active under different environmental conditions; the B.japonicum protein in this cluster is expressed constitutively; in Rhodobacter, Corynebacterium and Rhizobium this protein is essential for growth"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="groEL" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6607" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7122" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="16" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7149-7165" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="10" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="6" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7557-288" color="#ffff00" type="standard"/><Q name="note"><V text="ColE origin"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="5" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7123-7127" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="4" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7851" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="6910-6943" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="6910-6914" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="6915-6943" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6915-6943" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="6915-6943" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="1" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="6916-6943" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6916-6943" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6916-6943" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6916-6943" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="2" name="P224 SP_p1715_F" sequence="tatcttcttctctcatcattagaagag" description="0"><BindingSite location="6658-6684" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6658-6684" bases="tatcttcttctctcatcattagaagag"/></BindingSite><BindingSite simplified="1" location="6658-6684" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6658-6684" bases="tatcttcttctctcatcattagaagag"/></BindingSite></Primer><Primer recentID="3" name="P225 SP_p1715_R" sequence="tccattacaagatagagtattagttcg" description="0"><BindingSite location="6888-6914" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6888-6914" bases="tccattacaagatagagtattagttcg"/></BindingSite><BindingSite simplified="1" location="6888-6914" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6888-6914" bases="tccattacaagatagagtattagttcg"/></BindingSite></Primer><Primer recentID="4" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="7067-7093" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7067-7093" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="7067-7093" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7067-7093" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="5" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7176-7195" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7176-7195" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7176-7195" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7176-7195" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="6" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="15:11:41">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  I'               .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NASI6)4FIhj]Tky~M(,gFA!ӪGYc5'K8IMvSٮjMtg}(3=XjnUiR^j9")wۓjq:ۅYWکhiVQ5aEתz}j}]7תvBL=}t_9
6JQѪ7^Yn`qb]nVMcnuTS5.FhKUUZEj*-]NjiU^NV]	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%      !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  (M<?xml version="1.0"?><Features nextValidID="26"><Feature recentID="25" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7024-7024" color="#ffffff" type="standard"/></Feature><Feature recentID="24" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7031-7036" color="#a6acb3" type="standard"/></Feature><Feature recentID="23" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7052-7057" color="#a6acb3" type="standard"/></Feature><Feature recentID="22" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6887-7128" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="21" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6988-7058" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="20" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6665-6691" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="19" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6895-6921" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="18" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6923-6950" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="17" name="co-chaperonin GroES" directionality="2" translationMW="10272.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6645-6932" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="3" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6645-6932" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="chaperonin GroEL" directionality="2" translationMW="17231.43" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="5858" maxFusedRunOn="5858"><Segment range="6108-6607" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="60 kDa chaperone family; promotes refolding of misfolded polypeptides especially under stressful conditions; forms two stacked rings of heptamers to form a barrel-shaped 14mer; ends can be capped by GroES; misfolded proteins enter the barrel where they are refolded when GroES binds; many bacteria have multiple copies of the groEL gene which are active under different environmental conditions; the B.japonicum protein in this cluster is expressed constitutively; in Rhodobacter, Corynebacterium and Rhizobium this protein is essential for growth"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="groEL" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6607" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7128" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="16" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7155-7171" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="10" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="6" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7563-288" color="#ffff00" type="standard"/><Q name="note"><V text="ColE origin"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="5" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7129-7133" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="4" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7857" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="6916-6949" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="6916-6920" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6921-6949" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="1" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="2" name="P224 SP_p1715_F" sequence="tatcttcttctctcatcattagaagag" description="0"><BindingSite location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite><BindingSite simplified="1" location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite></Primer><Primer recentID="3" name="P225 SP_p1715_R" sequence="tccattacaagatagagtattagttcg" description="0"><BindingSite location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite><BindingSite simplified="1" location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite></Primer><Primer recentID="4" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="5" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7182-7201" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7182-7201" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7182-7201" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7182-7201" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="6" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="15:11:36">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  I%               .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NASI6)4FIhj]Tky~M(,gFA!ӪGYc5'K8IMvSٮjMtg}(3=XjnUiR^j9")wۓjq:ۅYWکjj%cYgmQ*}z&z}zod$#oZ}Mu~ikMq+e逸iڡj&6[J;RfUUVƩzUeПk%-"kgM"T>XHN^ݥ(?:,~,N63 D0ɠ3󯳷,8	PY\T]MT=5PU2?MUls-teF}N<gL:d$&+N|>      !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  (M<?xml version="1.0"?><Features nextValidID="26"><Feature recentID="25" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7018-7018" color="#ffffff" type="standard"/></Feature><Feature recentID="24" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7025-7030" color="#a6acb3" type="standard"/></Feature><Feature recentID="23" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7046-7051" color="#a6acb3" type="standard"/></Feature><Feature recentID="22" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6881-7122" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="21" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6982-7052" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="20" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6659-6685" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="19" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6889-6915" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="18" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6917-6944" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="17" name="co-chaperonin GroES" directionality="2" translationMW="10272.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6639-6926" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="3" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6639-6926" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="chaperonin GroEL" directionality="2" translationMW="17231.43" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="5858" maxFusedRunOn="5858"><Segment range="6108-6607" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="60 kDa chaperone family; promotes refolding of misfolded polypeptides especially under stressful conditions; forms two stacked rings of heptamers to form a barrel-shaped 14mer; ends can be capped by GroES; misfolded proteins enter the barrel where they are refolded when GroES binds; many bacteria have multiple copies of the groEL gene which are active under different environmental conditions; the B.japonicum protein in this cluster is expressed constitutively; in Rhodobacter, Corynebacterium and Rhizobium this protein is essential for growth"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="groEL" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6607" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7122" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="16" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7149-7165" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="10" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="6" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7557-288" color="#ffff00" type="standard"/><Q name="note"><V text="ColE origin"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="5" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7123-7127" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="4" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7851" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="6910-6943" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="6910-6914" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="6915-6943" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6915-6943" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="6915-6943" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="1" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="6916-6943" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6916-6943" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6916-6943" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6916-6943" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="2" name="P224 SP_p1715_F" sequence="tatcttcttctctcatcattagaagag" description="0"><BindingSite location="6658-6684" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6658-6684" bases="tatcttcttctctcatcattagaagag"/></BindingSite><BindingSite simplified="1" location="6658-6684" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6658-6684" bases="tatcttcttctctcatcattagaagag"/></BindingSite></Primer><Primer recentID="3" name="P225 SP_p1715_R" sequence="tccattacaagatagagtattagttcg" description="0"><BindingSite location="6888-6914" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6888-6914" bases="tccattacaagatagagtattagttcg"/></BindingSite><BindingSite simplified="1" location="6888-6914" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6888-6914" bases="tccattacaagatagagtattagttcg"/></BindingSite></Primer><Primer recentID="4" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="7067-7093" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7067-7093" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="7067-7093" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7067-7093" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="5" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7176-7195" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7176-7195" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7176-7195" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7176-7195" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="6" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="15:11:14">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  I&               .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
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ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
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6JQѪ7^Yn`qb]nVMcnuTS5.FhKUUZEj*-]NjiU^NV]	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%      !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  (M<?xml version="1.0"?><Features nextValidID="26"><Feature recentID="25" name="TSS" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7024-7024" color="#ffffff" type="standard"/></Feature><Feature recentID="24" name="GroES -10" directionality="2" type="-10_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7031-7036" color="#a6acb3" type="standard"/></Feature><Feature recentID="23" name="GroES -35" directionality="2" type="-35_signal" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="7052-7057" color="#a6acb3" type="standard"/></Feature><Feature recentID="22" name="pF promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6887-7128" color="#0080ff" type="standard"/><Q name="note"><V text="pF promoter"/></Q></Feature><Feature recentID="21" name="pF2 promoter" directionality="2" type="misc_feature" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6988-7058" color="#0080ff" type="standard"/><Q name="note"><V text="pF2 promoter"/></Q></Feature><Feature recentID="20" name="P224 SP_p1715_F" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6665-6691" color="#8080ff" type="standard"/><Q name="note"><V text="P224 SP_p1715_F"/></Q></Feature><Feature recentID="19" name="P225 SP_p1715_R" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6895-6921" color="#8080ff" type="standard"/><Q name="note"><V text="P225 SP_p1715_R"/></Q></Feature><Feature recentID="18" name="P051 groESpromoR" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6923-6950" color="#0080ff" type="standard"/><Q name="note"><V text="P051 groESpromoR"/></Q></Feature><Feature recentID="17" name="co-chaperonin GroES" directionality="2" translationMW="10272.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6645-6932" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="10 kDa chaperonin; Cpn10; GroES; forms homoheptameric ring; binds to one or both ends of the GroEL double barrel in the presence of adenine nucleotides capping it; folding of unfolded substrates initiates in a GroEL-substrate bound and capped by GroES; release of the folded substrate is dependent on ATP binding and hydrolysis in the trans ring"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="3" name="groES" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6645-6932" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="chaperonin GroEL" directionality="2" translationMW="17231.43" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="5858" maxFusedRunOn="5858"><Segment range="6108-6607" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="60 kDa chaperone family; promotes refolding of misfolded polypeptides especially under stressful conditions; forms two stacked rings of heptamers to form a barrel-shaped 14mer; ends can be capped by GroES; misfolded proteins enter the barrel where they are refolded when GroES binds; many bacteria have multiple copies of the groEL gene which are active under different environmental conditions; the B.japonicum protein in this cluster is expressed constitutively; in Rhodobacter, Corynebacterium and Rhizobium this protein is essential for growth"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="groEL" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6607" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7128" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="16" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7155-7171" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="10" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="6" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7563-288" color="#ffff00" type="standard"/><Q name="note"><V text="ColE origin"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="5" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7129-7133" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="4" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7857" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="6916-6949" boundStrand="0" annealedBases="atGGACgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="at"/><Component hybridizedRange="6916-6920" bases="atGGA"/><Component bases="TC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6921-6949" boundStrand="0" annealedBases="Cgaatgttcataacaatcttactcctttg" meltingTemperature="56"><Component bases="atatGGATC"/><Component hybridizedRange="6921-6949" bases="Cgaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="1" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="6922-6949" boundStrand="0" annealedBases="gaatgttcataacaatcttactcctttg" meltingTemperature="55"><Component bases="atatCTGCAG"/><Component hybridizedRange="6922-6949" bases="gaatgttcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="49"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="2" name="P224 SP_p1715_F" sequence="tatcttcttctctcatcattagaagag" description="0"><BindingSite location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite><BindingSite simplified="1" location="6664-6690" boundStrand="0" annealedBases="tatcttcttctctcatcattagaagag" meltingTemperature="53"><Component hybridizedRange="6664-6690" bases="tatcttcttctctcatcattagaagag"/></BindingSite></Primer><Primer recentID="3" name="P225 SP_p1715_R" sequence="tccattacaagatagagtattagttcg" description="0"><BindingSite location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite><BindingSite simplified="1" location="6894-6920" boundStrand="1" annealedBases="tccattacaagatagagtattagttcg" meltingTemperature="54"><Component hybridizedRange="6894-6920" bases="tccattacaagatagagtattagttcg"/></BindingSite></Primer><Primer recentID="4" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="7073-7099" boundStrand="0" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="7073-7099" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="56"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="5" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7182-7201" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7182-7201" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7182-7201" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7182-7201" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="6" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  A<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="15:11:5">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NA'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features recycledIDs="0,1,2,3" nextValidID="17"><Feature recentID="16" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6134-6150" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="10" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="6" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6542-288" color="#ffff00" type="standard"/><Q name="note"><V text="ColE origin"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="5" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6108-6112" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="4" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-6836" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="4,3,2,1,0" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="5" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6161-6180" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="6161-6180" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="6161-6180" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="6161-6180" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="6" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  A<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="15:0:43">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  7t       5        5.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
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n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAiY-DVf|Yi-a]Uөffedٵ)Y9fXXUo!ŽYVzHVik蚪b*InnubE&tz*IR8*ިeQiTUӞ^Rda^YE0oEQyJ)Ui5Y5&EIgTTgffmf_Q;eYRvUVUEi֤emUYI4XIVxVJZWهFDjeE	(F>gQYjYj	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%    
    [  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  i<?xml version="1.0"?><Features nextValidID="17"><Feature recentID="16" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7287-7303" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="15" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="10" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="6" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7695-288" color="#ffff00" type="standard"/><Q name="note"><V text="ColE origin"/></Q><Q name="direction"><V text="LEFT"/></Q></Feature><Feature recentID="5" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7261-7265" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="4" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7989" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="3" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="2" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6659-7260" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="1" name="rpsU" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6659-6685" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="30S ribosomal protein S21" directionality="2" translationMW="866.05" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6659-6685" color="#ff0000" type="standard" translated="1"/><Q name="note"><V text="a small basic protein that is one of the last in the subunit assembly; omission does not prevent assembly but the subunit is inactive; binds central domain of 16S rRNA"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="KROL128 ∆rpsU3 seq primer F" sequence="aacttgttttttaagtattttaagtgctg" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6531-6559" boundStrand="0" annealedBases="aacttgttttttaagtattttaagtgctg" meltingTemperature="54"><Component hybridizedRange="6531-6559" bases="aacttgttttttaagtattttaagtgctg"/></BindingSite><BindingSite simplified="1" location="6531-6559" boundStrand="0" annealedBases="aacttgttttttaagtattttaagtgctg" meltingTemperature="54"><Component hybridizedRange="6531-6559" bases="aacttgttttttaagtattttaagtgctg"/></BindingSite></Primer><Primer recentID="1" name="KROL127 ∆rpsU3 FR2 R" sequence="tttaatgaaaattaggtattttagttaaatc" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7229-7259" boundStrand="1" annealedBases="tttaatgaaaattaggtattttagttaaatc" meltingTemperature="50"><Component hybridizedRange="7229-7259" bases="tttaatgaaaattaggtattttagttaaatc"/></BindingSite><BindingSite simplified="1" location="7229-7259" boundStrand="1" annealedBases="tttaatgaaaattaggtattttagttaaatc" meltingTemperature="50"><Component hybridizedRange="7229-7259" bases="tttaatgaaaattaggtattttagttaaatc"/></BindingSite></Primer><Primer recentID="2" name="KROL126 ∆rpsU3 FR1 F" sequence="atctacctgattatttacttattttgataa" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6107-6136" boundStrand="0" annealedBases="atctacctgattatttacttattttgataa" meltingTemperature="51"><Component hybridizedRange="6107-6136" bases="atctacctgattatttacttattttgataa"/></BindingSite><BindingSite simplified="1" location="6107-6136" boundStrand="0" annealedBases="atctacctgattatttacttattttgataa" meltingTemperature="51"><Component hybridizedRange="6107-6136" bases="atctacctgattatttacttattttgataa"/></BindingSite></Primer><Primer recentID="3" name="KROL125 ∆rpsU3 FR2 F" sequence="atatGCGGCCGCtcttggcatattatcctctcttaatttt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6668-6703" boundStrand="0" annealedBases="GCGGCCGCtcttggcatattatcctctcttaatttt" meltingTemperature="66"><Component bases="atat"/><Component hybridizedRange="6668-6703" bases="GCGGCCGCtcttggcatattatcctctcttaatttt"/></BindingSite><BindingSite simplified="1" location="6668-6703" boundStrand="0" annealedBases="GCGGCCGCtcttggcatattatcctctcttaatttt" meltingTemperature="66"><Component bases="atat"/><Component hybridizedRange="6668-6703" bases="GCGGCCGCtcttggcatattatcctctcttaatttt"/></BindingSite></Primer><Primer recentID="4" name="KROL124 ∆rpsU3 FR1 R" sequence="atatGCGGCCGCTtcttattagttcaagtcataaagctatc" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6639-6675" boundStrand="1" annealedBases="GCGGCCGCTtcttattagttcaagtcataaagctatc" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="6639-6675" bases="GCGGCCGCTtcttattagttcaagtcataaagctatc"/></BindingSite><BindingSite simplified="1" location="6639-6675" boundStrand="1" annealedBases="GCGGCCGCTtcttattagttcaagtcataaagctatc" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="6639-6675" bases="GCGGCCGCTtcttattagttcaagtcataaagctatc"/></BindingSite></Primer><Primer recentID="5" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7314-7333" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7314-7333" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7314-7333" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7314-7333" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="6" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:56:29">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  52       6        6.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
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ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAiY-DVf|Yi-a]Uөffedٵ)Y9fXXUo!ŽYVzHVik蚪b*InnubE&tz*IR8*ިeQiTUӞ^Rda^YE0oEQyJ)Ui5Y5&EIgTTgffmf_Q;eYRvUVUEi֤emUYI4XIVxVJZWهFDjeE	(F>gQYjYjB|NŬضN*Vo;uw]O6TSa!9{v;x=8X[\£&ξ3L&ӷC	g
ot[@sAQv!5PC_%T<82Cq6UԴBWy2;#;+ϫ2퓷8"B8"7o	    
    \  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features recycledIDs="4,5,6" nextValidID="20"><Feature recentID="3" name="30S ribosomal protein S21" directionality="2" translationMW="866.05" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6659-6685" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="a small basic protein that is one of the last in the subunit assembly; omission does not prevent assembly but the subunit is inactive; binds central domain of 16S rRNA"/></Q></Feature><Feature recentID="2" name="rpsU" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6659-6685" color="#00e600" type="standard"/></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6659-7261" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="19" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7990" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="18" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7262-7266" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="17" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7696-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="16" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="15" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7288-7304" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  B<?xml version="1.0"?><Primers recycledIDs="6" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="5" name="KROL127 ∆rpsU3 FR2 R" sequence="ttttaatgaaaattaggtattttagttaaatc" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7229-7260" boundStrand="1" annealedBases="ttttaatgaaaattaggtattttagttaaatc" meltingTemperature="50"><Component hybridizedRange="7229-7260" bases="ttttaatgaaaattaggtattttagttaaatc"/></BindingSite><BindingSite simplified="1" location="7229-7260" boundStrand="1" annealedBases="ttttaatgaaaattaggtattttagttaaatc" meltingTemperature="50"><Component hybridizedRange="7229-7260" bases="ttttaatgaaaattaggtattttagttaaatc"/></BindingSite></Primer><Primer recentID="0" name="KROL126 ∆rpsU3 FR1 F" sequence="atctacctgattatttacttattttgataa" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6107-6136" boundStrand="0" annealedBases="atctacctgattatttacttattttgataa" meltingTemperature="51"><Component hybridizedRange="6107-6136" bases="atctacctgattatttacttattttgataa"/></BindingSite><BindingSite simplified="1" location="6107-6136" boundStrand="0" annealedBases="atctacctgattatttacttattttgataa" meltingTemperature="51"><Component hybridizedRange="6107-6136" bases="atctacctgattatttacttattttgataa"/></BindingSite></Primer><Primer recentID="1" name="KROL125 ∆rpsU3 FR2 F" sequence="atatGCGGCCGCtcttggcatattatcctctcttaatttt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6668-6703" boundStrand="0" annealedBases="GCGGCCGCtcttggcatattatcctctcttaatttt" meltingTemperature="66"><Component bases="atat"/><Component hybridizedRange="6668-6703" bases="GCGGCCGCtcttggcatattatcctctcttaatttt"/></BindingSite><BindingSite simplified="1" location="6668-6703" boundStrand="0" annealedBases="GCGGCCGCtcttggcatattatcctctcttaatttt" meltingTemperature="66"><Component bases="atat"/><Component hybridizedRange="6668-6703" bases="GCGGCCGCtcttggcatattatcctctcttaatttt"/></BindingSite></Primer><Primer recentID="2" name="KROL124 ∆rpsU3 FR1 R" sequence="atatGCGGCCGCTtcttattagttcaagtcataaagctatc" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6639-6675" boundStrand="1" annealedBases="GCGGCCGCTtcttattagttcaagtcataaagctatc" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="6639-6675" bases="GCGGCCGCTtcttattagttcaagtcataaagctatc"/></BindingSite><BindingSite simplified="1" location="6639-6675" boundStrand="1" annealedBases="GCGGCCGCTtcttattagttcaagtcataaagctatc" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="6639-6675" bases="GCGGCCGCTtcttattagttcaagtcataaagctatc"/></BindingSite></Primer><Primer recentID="3" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7315-7334" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7315-7334" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7315-7334" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7315-7334" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="4" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:43:57">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  +       5        5.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAiY-DVf|Yi-a]Uөffedٵ)Y9fXXUo!ŽYVzHVik蚪b*InnubE&tz*IR8*ިeQiTUӞ^Rda^YD6iiE+PVidTjD:{Y%RoQr]}FI(njYfXhoUIUYV)f_ZUUUe&NEa&iYZ`M*mjY_fXe$TjFdefQgU	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%    
    [  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features recycledIDs="4,5,6" nextValidID="20"><Feature recentID="3" name="30S ribosomal protein S21" directionality="2" translationMW="842.11" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="6598" maxFusedRunOn="6598"><Segment range="6659-6684" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="a small basic protein that is one of the last in the subunit assembly; omission does not prevent assembly but the subunit is inactive; binds central domain of 16S rRNA"/></Q></Feature><Feature recentID="2" name="rpsU" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6659-6684" color="#00e600" type="standard"/></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6659-7260" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="19" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7989" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="18" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7261-7265" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="17" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7695-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="16" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="15" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7287-7303" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="2,1,0,5,6" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="3" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7314-7333" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7314-7333" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7314-7333" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7314-7333" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="4" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:18:33">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  *       -        -.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAiY-DVf|Yi-a]Uөffedٵ)Y9fXXUo!ŽYVzHVik蚪b*InnubE&tz*IR8*ިeQiTUӞ^Rda^YF6iiE+PVidTjD:{Y%RoQr]}FI(njYfXhoUIUYV)f_ZUUUe&NEa&iYZ`M*mjY_fXe$TjFdefQgU	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%    S  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features recycledIDs="4,5,6" nextValidID="20"><Feature recentID="3" name="30S ribosomal protein S21" directionality="2" translationMW="652.78" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6659-6676" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="a small basic protein that is one of the last in the subunit assembly; omission does not prevent assembly but the subunit is inactive; binds central domain of 16S rRNA"/></Q></Feature><Feature recentID="2" name="rpsU" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6659-6676" color="#00e600" type="standard"/></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6659-7252" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="19" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7981" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="18" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7253-7257" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="17" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7687-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="16" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="15" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7279-7295" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="2,1,0,5,6" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="3" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7306-7325" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7306-7325" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7306-7325" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7306-7325" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="4" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:16:18">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  *       0        0.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAiY-DVf|Yi-a]Uөffedٵ)Y9fXXUo!ŽYVzHVik蚪b*InnubE&tz*IR8*ިeQiTUӞ^Rda^YE٦ZYDBYUYRnTQjyduIFvivfecj&XeaU'khjUeXVa}jJQVQVU;SMgi4e}tdFaT[l`Rjc~UQFj^UiT'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ    V  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features recycledIDs="4,5,6" nextValidID="20"><Feature recentID="3" name="30S ribosomal protein S21" directionality="2" translationMW="815.96" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6659-6679" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="a small basic protein that is one of the last in the subunit assembly; omission does not prevent assembly but the subunit is inactive; binds central domain of 16S rRNA"/></Q></Feature><Feature recentID="2" name="rpsU" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6659-6679" color="#00e600" type="standard"/></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6659-7255" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="19" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7984" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="18" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7256-7260" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="17" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7690-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="16" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="15" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7282-7298" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="2,1,0,5,6" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="3" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7309-7328" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7309-7328" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7309-7328" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7309-7328" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="4" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:15:22">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  +               .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAiY-DVf|Yi-a]Uөffedٵ)Y9fXXUo!ŽYVzHVik蚪b*InnubE&tz*IR8*ިeQiTUӞ^Rda^YE~:>Y49"cJg㪷SZGmT*ךk٦ZYDBYUYRnTQjyduIFvivfecj&XeaU'khjUeXVa}jJQVQVU;SMgi4e}tdFaT[l`Rjc~UQFj^UiT'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ    
  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features recycledIDs="4,5,6" nextValidID="20"><Feature recentID="3" name="30S ribosomal protein S21" directionality="2" translationMW="7816.44" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6659-6859" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="a small basic protein that is one of the last in the subunit assembly; omission does not prevent assembly but the subunit is inactive; binds central domain of 16S rRNA"/></Q></Feature><Feature recentID="2" name="rpsU" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6659-6859" color="#00e600" type="standard"/></Feature><Feature recentID="1" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6659-7435" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="19" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-8164" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="18" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7436-7440" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="17" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7870-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="16" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="15" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7462-7478" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="2,1,0,5,6" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="3" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7489-7508" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7489-7508" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7489-7508" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7489-7508" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="4" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  A<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:14:7">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  &     Q          ۛ.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAiY-DVf|Yi-a]Uөffedٵ)Y9fXXUo!ŽYVzHVik蚪b*InnubE&tz*IR8*ިeQiTUӞ^RdaPk%-"kgM"T>XHN^ݥ(?:,~,N63 D0ɠ3󯳷,8	PY\T]MT=5PU2?MUls-teF}N<gL:d$&+N|>      !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  2<?xml version="1.0"?><Features recycledIDs="1,2,3,4,5,6" nextValidID="20"><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="19" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7387" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="18" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6659-6663" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="17" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7093-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="16" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="15" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6685-6701" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="2,1,0,5,6" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="3" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6712-6731" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="6712-6731" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="6712-6731" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="6712-6731" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="4" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:12:52">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  $c   
             .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NA'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  #<?xml version="1.0"?><Features recycledIDs="0,1,2,3,4,5,6" nextValidID="20"><Feature recentID="19" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-6836" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="18" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6108-6112" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="17" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6542-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="16" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="15" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6134-6150" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="2,1,0,5,6" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="3" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6161-6180" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="6161-6180" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="6161-6180" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="6161-6180" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="4" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:11:51">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  :   	            ϛ.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAU%T5Ui'-d6ei(J9ZDFfy̑cRY<WW_fVT[|FY6VtIn[`J_i:6gbeYK%rY^LBc<PklUujaQ-aUUZɃYUMUYѷZ8UFMYBTDdVh{YMyXaD(+EdKlaQ	V5"UT^	Fa5ILV:Pk%-"kgM"T>XHN^ݥ(?:,~,N63 D0ɠ3󯳷,8	PY\T]MT=5PU2?MUls-teF}N<gL:d$&+N|>          !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features nextValidID="20"><Feature recentID="19" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7887" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="18" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7159-7163" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="17" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7593-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="16" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="15" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="14" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="13" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="11" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7185-7201" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7158" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="5" name="FTL_1046" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6369" color="#00e600" type="standard"/></Feature><Feature recentID="4" name="D-alanyl-D-alanine carboxypeptidase" directionality="2" translationMW="9685.55" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="6097" maxFusedRunOn="6097"><Segment range="6108-6369" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature><Feature recentID="3" name="rpsU" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6625-6651" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="30S ribosomal protein S21" directionality="1" translationMW="831.04" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6625-6651" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="a small basic protein that is one of the last in the subunit assembly; omission does not prevent assembly but the subunit is inactive; binds central domain of 16S rRNA"/></Q></Feature><Feature recentID="1" name="FTL_1048" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6712-7158" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="hypothetical protein" directionality="1" translationMW="16767.04" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6712-7158" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q></Feature></Features>
  t<?xml version="1.0"?><Primers nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="6" name="KROL123 ∆rpsU2 FR2R" sequence="ttaagctaagttagatttcacaatagcg" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7130-7157" boundStrand="1" annealedBases="ttaagctaagttagatttcacaatagcg" meltingTemperature="55"><Component hybridizedRange="7130-7157" bases="ttaagctaagttagatttcacaatagcg"/></BindingSite><BindingSite simplified="1" location="7130-7157" boundStrand="1" annealedBases="ttaagctaagttagatttcacaatagcg" meltingTemperature="55"><Component hybridizedRange="7130-7157" bases="ttaagctaagttagatttcacaatagcg"/></BindingSite></Primer><Primer recentID="5" name="KROL122 ∆rpsU2 FR1F" sequence="taaaattactaggtatcgtaaaaagtgc" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6107-6134" boundStrand="0" annealedBases="taaaattactaggtatcgtaaaaagtgc" meltingTemperature="53"><Component hybridizedRange="6107-6134" bases="taaaattactaggtatcgtaaaaagtgc"/></BindingSite><BindingSite simplified="1" location="6107-6134" boundStrand="0" annealedBases="taaaattactaggtatcgtaaaaagtgc" meltingTemperature="53"><Component hybridizedRange="6107-6134" bases="taaaattactaggtatcgtaaaaagtgc"/></BindingSite></Primer><Primer recentID="0" name="KROL121 ∆rpsU2 seq primer F" sequence="ttagaataatccactaattgaaatattgg" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6531-6559" boundStrand="0" annealedBases="ttagaataatccactaattgaaatattgg" meltingTemperature="52"><Component hybridizedRange="6531-6559" bases="ttagaataatccactaattgaaatattgg"/></BindingSite><BindingSite simplified="1" location="6531-6559" boundStrand="0" annealedBases="ttagaataatccactaattgaaatattgg" meltingTemperature="52"><Component hybridizedRange="6531-6559" bases="ttagaataatccactaattgaaatattgg"/></BindingSite></Primer><Primer recentID="1" name="KROL119 ∆rpsU2 FR2F" sequence="atatGCGGCCGCTaaaaggtagttaaatttaattcatcgttg" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6633-6670" boundStrand="0" annealedBases="GCGGCCGCTaaaaggtagttaaatttaattcatcgttg" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="6633-6670" bases="GCGGCCGCTaaaaggtagttaaatttaattcatcgttg"/></BindingSite><BindingSite simplified="1" location="6633-6670" boundStrand="0" annealedBases="GCGGCCGCTaaaaggtagttaaatttaattcatcgttg" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="6633-6670" bases="GCGGCCGCTaaaaggtagttaaatttaattcatcgttg"/></BindingSite></Primer><Primer recentID="2" name="KROL120 ∆rpsU2 FR1R" sequence="atatGCGGCCGCgcttggcataccctctcacctcg" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6610-6640" boundStrand="1" annealedBases="GCGGCCGCgcttggcataccctctcacctcg" meltingTemperature="74"><Component bases="atat"/><Component hybridizedRange="6610-6640" bases="GCGGCCGCgcttggcataccctctcacctcg"/></BindingSite><BindingSite simplified="1" location="6610-6640" boundStrand="1" annealedBases="GCGGCCGCgcttggcataccctctcacctcg" meltingTemperature="74"><Component bases="atat"/><Component hybridizedRange="6610-6640" bases="GCGGCCGCgcttggcataccctctcacctcg"/></BindingSite></Primer><Primer recentID="3" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7212-7231" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7212-7231" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7212-7231" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7212-7231" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="4" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  B<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:11:40">2019.2.8</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  .)               ƛ.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
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ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAU%T5Ui'-d6ei(J9ZDFfy̑cRY<WW_fVT[|FY6VtIn[`J_i:6gbeYK%rY^LBcBJVJPZV-ժ҇DVTeVF]k&fWU4haUgFBhZuVSTuچv5dNUQYgc5ad9.(D`%[`ԉUQz8$%3![B|NŬضN*Vo;uw]O6TSa!9{v;x=8X[\£&ξ3L&ӷC	g
ot[@sAQv!5PC_%T<82Cq6UԴBWy2;#;+ϫ2퓷8"B8"7o	      !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
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omission does not prevent assembly but the subunit is inactive; binds central domain of 16S rRNA"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="3" name="rpsU" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6625-6642" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="D-alanyl-D-alanine carboxypeptidase" directionality="2" translationMW="9685.55" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="6097" maxFusedRunOn="6097"><Segment range="6108-6369" color="#ff0000" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="FTL_1046" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6369" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7149" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="24" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7176-7192" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="23" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="18" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="16" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="15" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="14" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7584-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="13" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7150-7154" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="10" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7878" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="6,5,4,3,0" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="1" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7203-7222" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7203-7222" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7203-7222" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7203-7222" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="2" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  A<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="23:10:3">2019.2.7</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  .W       z        z.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAU%T5Ui'-d6ei(J9ZDFfy̑cRY<WW_fVT[|FY6VtIn[`J_i:6gbeYK%rY^LBcEED_EFPY&y$ETTUU)TNDeBJVJPZV-ժ҇DVTeVF]k&fWU4haUgFBhZuVSTuچv5dNUQYgc5ad9.(D`%[`ԉUQz8$%3![B|NŬضN*Vo;uw]O6TSa!9{v;x=8X[\£&ξ3L&ӷC	g
ot[@sAQv!5PC_%T<82Cq6UԴBWy2;#;+ϫ2퓷8"B8"7o	      !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features recycledIDs="7,8,9,11,12" nextValidID="25"><Feature recentID="6" name="hypothetical protein" directionality="1" translationMW="16767.04" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6883-7329" color="#ff0000" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="5" name="FTL_1048" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6883-7329" color="#00e600" type="standard"/></Feature><Feature recentID="4" name="30S ribosomal protein S21" directionality="1" translationMW="7840.48" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="6625-6822" color="#ff0000" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="a small basic protein that is one of the last in the subunit assembly; omission does not prevent assembly but the subunit is inactive; binds central domain of 16S rRNA"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="3" name="rpsU" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6625-6822" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="D-alanyl-D-alanine carboxypeptidase" directionality="2" translationMW="9685.55" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="6097" maxFusedRunOn="6097"><Segment range="6108-6369" color="#ff0000" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="FTL_1046" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6108-6369" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7329" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="24" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7356-7372" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="23" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="18" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="16" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="15" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="14" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7764-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="13" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7330-7334" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="10" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-8058" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="6,5,4,3,0" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="1" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7383-7402" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7383-7402" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7383-7402" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7383-7402" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="2" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  A<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="23:9:25">2019.2.7</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  $p                .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NA'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  2<?xml version="1.0"?><Features recycledIDs="0,1,2,3,4,5,6,7,8,9,11,12" nextValidID="25"><Feature recentID="24" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6134-6150" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="23" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="18" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="16" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="15" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="14" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6542-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="13" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6108-6112" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="10" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-6836" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers recycledIDs="6,5,4,3,0" nextValidID="7"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="1" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6161-6180" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="6161-6180" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="6161-6180" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="6161-6180" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="2" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  @<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="23:5:0">2019.2.7</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  B       B        B.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAW~ZjJUnڹhV;aU
^YYh0Z[>ީ.ZUi*+Ӛn:vz)OJ	TcbTkzuZ9_~kLvwPRmkN5I5im9S'JiMXj^jKE^IbFni3ij剕%|juN|NŬضN*Vo;uw]O6TSa!9{v;x=8X[\£&ξ3L&ӷC	g
ot[@sAQv!5PC_%T<82Cq6UԴBWy2;#;+ϫ2퓷8"B8"7o	        h  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  %<?xml version="1.0"?><Features nextValidID="25"><Feature recentID="24" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7044-7060" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="23" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="22" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="21" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="20" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="19" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="18" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="17" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="16" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="15" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="14" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7452-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="13" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7018-7022" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q></Feature><Feature recentID="12" name="FTL_0880" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6293-6297" color="#a6acb3" type="standard"/><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="db_xref"><V text="3952354" predef="GeneID"/></Q></Feature><Feature recentID="11" name="FTL_0880" directionality="1" translationMW="75.07" type="CDS" allowSegmentOverlaps="0" readingFrame="3" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="3" maxRunOn="6306" maxFusedRunOn="6306"><Segment range="6293-6297" color="#993366" type="standard" translated="1"/><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q></Feature><Feature recentID="10" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-7746" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="9" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6302-7017" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="8" name="FTL_1251" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6302-6310" color="#00e600" type="standard"/></Feature><Feature recentID="7" name="proton-dependent oligopeptide transport" directionality="1" translationMW="246.24" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="512" hitsStopCodon="1"><Segment range="6302-6310" color="#ff0000" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="6" name="FTL_1252" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6307-7017" color="#00e600" type="standard"/></Feature><Feature recentID="5" name="ATP-dependent RNA helicase RhlE" 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name="transposase" directionality="1" translationMW="3618.53" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="193" hitsStopCodon="1"><Segment range="6108-6197" color="#ff0000" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="1" name="isftu2" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6197" color="#00e600" type="standard"/></Feature><Feature recentID="0" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6292" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features>
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  A<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="23:4:37">2019.2.7</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
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ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAUf[涮Z**j~UBhVAZ!<ꏨ*wj&mE
(4曥ljN"d'iS뒾UmbǭޝDVW=+demF5ZfMyGjfF[NTIҡ}y=6Wzi!LZbejIn,vӆpk%-"kgM"T>XHN^ݥ(?:,~,N63 D0ɠ3󯳷,8	PY\T]MT=5PU2?MUls-teF}N<gL:d$&+N|>    `  j  5  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
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name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="4" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="0" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6993-7009" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  <?xml version="1.0"?><Primers nextValidID="6"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7020-7039" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7020-7039" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7020-7039" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="54"><Component hybridizedRange="7020-7039" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="1" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer><Primer recentID="2" name="KROL3 DFTL1251_FR2F" sequence="atatGCGGCCGCCaataattaatcttttgcctttggttttct" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6240-6278" boundStrand="0" annealedBases="tGCGGCCGCCaataattaatcttttgcctttggttttct" meltingTemperature="67"><Component bases="ata"/><Component hybridizedRange="6240-6278" bases="tGCGGCCGCCaataattaatcttttgcctttggttttct"/></BindingSite><BindingSite simplified="1" location="6240-6278" boundStrand="0" annealedBases="tGCGGCCGCCaataattaatcttttgcctttggttttct" meltingTemperature="67"><Component bases="ata"/><Component hybridizedRange="6240-6278" bases="tGCGGCCGCCaataattaatcttttgcctttggttttct"/></BindingSite></Primer><Primer recentID="3" name="KROL4 DFTL1251_FR2R" sequence="atatGGTACCgtatcagctgatgttgtaaatacgac" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6940-6971" boundStrand="1" annealedBases="GGTACCgtatcagctgatgttgtaaatacgac" meltingTemperature="61"><Component bases="atat"/><Component hybridizedRange="6940-6971" bases="GGTACCgtatcagctgatgttgtaaatacgac"/></BindingSite><BindingSite simplified="1" location="6940-6971" boundStrand="1" annealedBases="GGTACCgtatcagctgatgttgtaaatacgac" meltingTemperature="61"><Component bases="atat"/><Component hybridizedRange="6940-6971" bases="GGTACCgtatcagctgatgttgtaaatacgac"/></BindingSite></Primer><Primer recentID="4" name="KROL1 DFTL1251_FR1F" sequence="atatGGATCCttagttttcaaaaatataatctttgctattc" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6101-6137" boundStrand="0" annealedBases="GGATCCttagttttcaaaaatataatctttgctattc" meltingTemperature="57"><Component bases="atat"/><Component hybridizedRange="6101-6137" bases="GGATCCttagttttcaaaaatataatctttgctattc"/></BindingSite><BindingSite simplified="1" location="6101-6137" boundStrand="0" annealedBases="GGATCCttagttttcaaaaatataatctttgctattc" meltingTemperature="57"><Component bases="atat"/><Component hybridizedRange="6101-6137" bases="GGATCCttagttttcaaaaatataatctttgctattc"/></BindingSite></Primer><Primer recentID="5" name="KROL2 DFTL1251_FR1R" sequence="atatGCGGCCGCaatagtcacaattattctcctattttaaac" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6211-6248" boundStrand="1" annealedBases="GCGGCCGCaatagtcacaattattctcctattttaaac" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="6211-6248" bases="GCGGCCGCaatagtcacaattattctcctattttaaac"/></BindingSite><BindingSite simplified="1" location="6211-6248" boundStrand="1" annealedBases="GCGGCCGCaatagtcacaattattctcctattttaaac" meltingTemperature="64"><Component bases="atat"/><Component hybridizedRange="6211-6248" bases="GCGGCCGCaatagtcacaattattctcctattttaaac"/></BindingSite></Primer></Primers>
  D<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="16:10:19">2018.10.23</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  1       C        C.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAUf[涮Z**j~UBhVAZ!<k%-"kgM"T>XHN^ݥ(?:,~,N63 D0ɠ3󯳷,8	PY\T]MT=5PU2?MUls-teF}N<gL:d$&+N|>    `  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  #*<?xml version="1.0"?><Features recycledIDs="11,12,13,14" nextValidID="25"><Feature recentID="24" name="proton-dependent oligopeptide transport" directionality="1" translationMW="331.43" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="6283" maxFusedRunOn="6283"><Segment range="6233-6241" color="#ff0000" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="23" name="FTL_1251" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6233-6241" color="#00e600" type="standard"/></Feature><Feature recentID="22" name="transposase" directionality="1" translationMW="1461.85" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="210" hitsStopCodon="1"><Segment range="6108-6146" color="#ff0000" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="codon_start"><V int="1"/></Q></Feature><Feature recentID="18" name="isftu2" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6146" color="#00e600" type="standard"/></Feature><Feature recentID="17" name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-6241" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="21" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-6979" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="20" name="FTL_0880" directionality="1" translationMW="75.07" type="CDS" allowSegmentOverlaps="0" readingFrame="3" consecutiveTranslationNumbering="1" consecutiveNumberingStartsFrom="3" maxRunOn="6354" maxFusedRunOn="6354"><Segment range="6242-6246" color="#993366" type="standard" translated="1"/><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q></Feature><Feature recentID="19" name="FTL_0880" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6242-6246" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952354" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q></Feature><Feature recentID="16" name="FTL_0882" directionality="2" translateFirstCodonAsMet="1" translationMW="117.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="6214" maxFusedRunOn="6214"><Segment range="6251-6255" color="#993366" type="standard" translated="1"/><Q name="db_xref"><V text="89256240" predef="GI"/><V text="3952356" predef="GeneID"/></Q><Q name="protein_id"><V text="YP_513602.1"/></Q><Q name="inference"><V bool="1" text="4.1" predef="profile"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="apolipoprotein N-acyltransferase"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="note"><V text="/dnas_title=apolipoprotein N-acyltransferase"/></Q></Feature><Feature recentID="15" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6251-6255" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="10" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6685-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="9" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="4" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="0" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6277-6293" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  Z<?xml version="1.0"?><Primers nextValidID="2"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="1" name="KROL1 DFTL1251_FR1F" sequence="ttagttttcaaaaatataatctttgctattc" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6107-6137" boundStrand="0" annealedBases="ttagttttcaaaaatataatctttgctattc" meltingTemperature="51"><Component hybridizedRange="6107-6137" bases="ttagttttcaaaaatataatctttgctattc"/></BindingSite><BindingSite simplified="1" location="6107-6137" boundStrand="0" annealedBases="ttagttttcaaaaatataatctttgctattc" meltingTemperature="51"><Component hybridizedRange="6107-6137" bases="ttagttttcaaaaatataatctttgctattc"/></BindingSite></Primer><Primer recentID="0" name="KROL2 DFTL1251_FR1R" sequence="aatagtcacaattattctcctattttaaac" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6211-6240" boundStrand="1" annealedBases="aatagtcacaattattctcctattttaaac" meltingTemperature="53"><Component hybridizedRange="6211-6240" bases="aatagtcacaattattctcctattttaaac"/></BindingSite><BindingSite simplified="1" location="6211-6240" boundStrand="1" annealedBases="aatagtcacaattattctcctattttaaac" meltingTemperature="53"><Component hybridizedRange="6211-6240" bases="aatagtcacaattattctcctattttaaac"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="21:35:2">2018.10.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  ;       ;        ;.	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAUf[涮Z**j~UBhVAZ!<:hkYz]Tj:U{ZUnkfiTjJDQNNIj=hrH`iQffiښWݚdڄ iUUE[ign֡%6YYi[SYFid(dij數暊*)Vd]WZaayYZeRjYVeVijtiiiV&Yi?[9VmugkoVI[jVfʎԋ7o$e@k%-"kgM"T>XHN^ݥ(?:,~,N63 D0ɠ3󯳷,8	PY\T]MT=5PU2?MUls-teF}N<gL:d$&+N|>    `  j  a  !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
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text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="4" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature 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  D<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="21:34:17">2018.10.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  1                .	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAϣ	XUFnYV)jiifJ^En։g*DMnhiv}e}٦II%X	UZd[hyZ?Fz
j[]QYnjEEU5JDfIbHZibnVIYUۍzoIUNVuU&fZ5ijjGJVeijfY=^Xv}fdeflHq&BJYji]4	;[b8!Z)ֽvq<R%O#億Z2Wial=s0C
;:;x8Ì10N%)m eE؄CUA|P(ʫuV;R1_ATgʿΟ~/>3NBnҰ+;8_+O3%      !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
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text="3952354" predef="GeneID"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q></Feature><Feature recentID="13" name="FTL_0880" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6122-6130" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952354" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q></Feature><Feature recentID="12" name="FTL_0881" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="6402" 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swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7567-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="9" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="4" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="0" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7159-7175" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
   <?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  D<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="21:34:13">2018.10.22</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  6        "        ".	Gq4]D&$0tJR>^py]&8SҟQ($	0SMs4UTon^ܥamU~FiU*Rfa%z[P(*3ktQhh;ֺ<:,Bý04u	0ϝoc6V!8%LoN/n~>M\<Qm P2NUif)9*ZT[:*U]UZj{YDFխV:WUeuA%)k\rCiZᩘ	X5#nlh?D5BLFx8pZmMo'c`1UkКEo!Vh+<(ƾ*%2j˳3[qae HjLRTV6W=!骇id(q,!*i\:Uf	fV*چ;Jh)h(x4M8z:Vѷ64| 5R[}x/~u;546pa6F>2ZeyiSiM
ha0gfbjDVWV@+ݫU#߇.Q_ivUd2m?.ʰ!WӿpNLGS3-0",0DѢxD[ꌪ^)htb43W(hfNmh΄'LR!%%BmF]=*EWe6%fG%-lџj<Tr2n))
ު*N<<ΣE*pQad)YQ$J􊎵g>뜒n2`τJ>'jr-Q+TN.ᜥ(%V$Y\F%b"ڍj,TsL8JǪEؕj9cTq$*1<ʢX=8W/CFjfڪ+@j4ٶfXb`ꂲ΄>0T)g:*^.bxL.:FJJsuUf% =Rgzwj>:Iu3{hׂdUUGV⮫kd6eJ>[uffyYdBf!TNDMlm|#0,J0=N/qj?M	>lPwd 8
n)$4M(MЌY@t;0 >zߛ(PT_V1UgUa)0U3*n|xՍix@DqTNwĀoS<2C(0!Ytc%U6͚Y]TJUo-:Z>*`:ro;{A+|uuѕqP.xO*>+̀wl32넬6  |xծ*Gθn&Vo=WOSkC׊@0>9=81PPƔ%@-(q=NAbfV9jefId)jfbeIilߖV*_J^Yx4mM2JRfD}rM%MĩnjnQO6mZ7זUzwYHdfJH$c֪٨jQmhA4gXeJ%ͦ>NWWYIeRFjE6Jg^ڢ%qJaiNa^NvvV_c*Ine.ꑎej훦 &&Zh'j,QUdYa)]deX9>&-cV^UNf^֦[ZjٚDU)zSZOZ%+i6Yif&&j`&UimhjAh꛸*uEmuFIfMmZjVyTy)Y%"ZnڪjyUY%eWn6ꩽ%V:eXkEVTUjdեZje*ZcEZiIZOg]dyaՒaڕ"ě	+e=;Yt'Zmd⯈hZuSeH> ih3ίK_#Ӎ*2h;+0m;{T0pE4bUMT=UC̣*d;m[KD QC*S3:|S ή;s	J$,S#q?8Ϧ          !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  *<?xml version="1.0"?><Features nextValidID="22"><Feature recentID="21" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-8888" color="#ffffff" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q></Feature><Feature recentID="20" name="FTL_0880" directionality="1" translateFirstCodonAsMet="1" translationMW="341.40" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="7238" maxFusedRunOn="7238"><Segment range="7131-7139" color="#993366" type="standard" translated="1"/><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q></Feature><Feature recentID="19" name="FTL_0880" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7131-7139" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952354" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q></Feature><Feature recentID="18" name="FTL_0879" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="6097" maxFusedRunOn="6097"><Segment range="6113-6702" color="#993366" type="standard" translated="1"/><Q name="db_xref"><V text="89256237" predef="GI"/><V text="3952353" predef="GeneID"/></Q><Q name="protein_id"><V text="YP_513599.1"/></Q><Q name="inference"><V bool="1" text="2004" predef="profile"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="beta-lactamase"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="FTL_0879"/></Q><Q name="note"><V text="/dnas_title=beta-lactamase"/></Q><Q name="EC_number"><V text="3.5.2.6"/></Q></Feature><Feature recentID="17" name="FTL_0879" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="6113-6702" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952353" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0879"/></Q></Feature><Feature recentID="16" name="FTL_0882" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="7943" maxFusedRunOn="7943"><Segment range="7974-8164" color="#993366" type="standard" translated="1"/><Q name="db_xref"><V text="89256240" predef="GI"/><V text="3952356" predef="GeneID"/></Q><Q name="protein_id"><V text="YP_513602.1"/></Q><Q name="inference"><V bool="1" text="4.1" predef="profile"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="apolipoprotein N-acyltransferase"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="note"><V text="/dnas_title=apolipoprotein N-acyltransferase"/></Q></Feature><Feature recentID="15" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7974-8164" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="14" name="FTL_0880" directionality="1" translateFirstCodonAsMet="1" translationMW="319.42" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="7238" maxFusedRunOn="7238"><Segment range="7149-7157" color="#993366" type="standard" translated="1"/><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q></Feature><Feature recentID="13" name="FTL_0880" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7149-7157" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952354" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q></Feature><Feature recentID="12" name="FTL_0881" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1" maxRunOn="7429" maxFusedRunOn="7429"><Segment range="7453-7728" color="#993366" type="standard" translated="1"/><Q name="db_xref"><V text="89256239" predef="GI"/><V text="3952355" predef="GeneID"/></Q><Q name="protein_id"><V text="YP_513601.1"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="locus_tag"><V text="FTL_0881"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q></Feature><Feature recentID="11" name="FTL_0881" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7453-7728" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952355" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0881"/></Q></Feature><Feature recentID="10" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8594-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="9" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="8" name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="7" name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="transl_table"><V int="1"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="4" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="3" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="2" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="1" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="0" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="8186-8202" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
   <?xml version="1.0"?><Primers nextValidID="0"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="23:40:14">2018.2.27</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
        ͮZTR

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text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7762-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6108-7327" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="6696..6701" colors="white"/></TopStrand><BottomStrand><ColorRange range="6696..6701" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..7326" colors="red"/></TopStrand><BottomStrand><ColorRange range="6107..7326" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6107"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="6836" ID="39" circular="1" resurrectable="1" operation="remove"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6134-6150" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6542-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..124" colors="blue-black"/><ColorRange range="5253..6106" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6107..124" colors="blue-black"/><ColorRange range="5253..6106" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6107" val2="8946"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="9676" ID="38" circular="1" resurrectable="1" operation="replace" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8974-8990" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9382-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="8337-8947" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="8947..479" colors="blue-black"/><ColorRange range="4898..6106" colors="black-red"/><ColorRange range="6107..8946" colors="white"/></TopStrand><BottomStrand><ColorRange range="8947..479" colors="blue-black"/><ColorRange range="4898..6106" colors="black-red"/><ColorRange range="6107..8946" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="8301..8306" colors="red"/></TopStrand><BottomStrand><ColorRange range="8301..8306" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="replace" val1="8301" val2="8306"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="9676" ID="37" circular="1" resurrectable="1" operation="replace" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8974-8990" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9382-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="8337-8947" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="8301..8306" colors="white"/></TopStrand><BottomStrand><ColorRange range="8301..8306" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="8301..8306" colors="red"/></TopStrand><BottomStrand><ColorRange range="8301..8306" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="replace" val1="8301" val2="8306"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="9676" ID="36" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8974-8990" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9382-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="8337-8947" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="8301..8306" colors="white"/></TopStrand><BottomStrand><ColorRange range="8301..8306" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="8301..9509" colors="blue-black"/><ColorRange range="7092..8300" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="8301..9509" colors="blue-black"/><ColorRange range="7092..8300" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="8301" val2="8301"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="9677" ID="35" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8975-8991" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9383-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="8338-8948" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-8337" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="8302..9510" colors="blue-black"/><ColorRange range="7092..8300" colors="black-red"/><ColorRange range="8301..8301" colors="white"/></TopStrand><BottomStrand><ColorRange range="8302..9510" colors="blue-black"/><ColorRange range="7092..8300" colors="black-red"/><ColorRange range="8301..8301" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="8301..9509" colors="blue-black"/><ColorRange range="7092..8300" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="8301..9509" colors="blue-black"/><ColorRange range="7092..8300" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="8301" val2="8302"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="9679" ID="34" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8977-8993" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9385-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="8340-8950" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-8339" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="8303..9511" colors="blue-black"/><ColorRange range="7092..8300" colors="black-red"/><ColorRange range="8301..8302" colors="white"/></TopStrand><BottomStrand><ColorRange range="8303..9511" colors="blue-black"/><ColorRange range="7092..8300" colors="black-red"/><ColorRange range="8301..8302" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="8303..8303" colors="red"/></TopStrand><BottomStrand><ColorRange range="8303..8303" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="8303"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="9678" ID="33" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-8338" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="8339-8949" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9384-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8976-8992" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="8301..8302" colors="red"/></TopStrand><BottomStrand><ColorRange range="8301..8302" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="8301"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="9676" ID="32" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-8336" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="8337-8947" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="9382-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8974-8990" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="8301..8306" colors="red"/></TopStrand><BottomStrand><ColorRange range="8301..8306" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="8301"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="9670" ID="31" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-8330" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in 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text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" 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gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8991-9007" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="8964..481" colors="blue-black"/><ColorRange range="7125..8335" colors="black-red"/><ColorRange range="8336..8963" colors="white"/></TopStrand><BottomStrand><ColorRange range="8964..481" colors="blue-black"/><ColorRange range="7125..8335" colors="black-red"/><ColorRange range="8336..8963" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..8335" colors="red"/></TopStrand><BottomStrand><ColorRange range="6107..8335" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6107"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="7464" ID="27" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7170-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6590" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6591-6735" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6762-6778" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..7039" colors="blue-black"/><ColorRange range="5174..6106" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6107..7039" colors="blue-black"/><ColorRange range="5174..6106" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6107" val2="6641"/><Node name="pKR35 pEX catalase mod rbs.dna" type="DNA" seqLen="7999" ID="26" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7125" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7705-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="7126-7270" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7297-7313" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="6642..7640" colors="blue-black"/><ColorRange range="5108..6106" colors="black-red"/><ColorRange range="6107..6641" colors="white"/></TopStrand><BottomStrand><ColorRange range="6642..7640" colors="blue-black"/><ColorRange range="5108..6106" colors="black-red"/><ColorRange range="6107..6641" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="7125..7269" colors="red"/></TopStrand><BottomStrand><ColorRange range="7125..7269" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="7125"/><Node name="pKR13 pEX GroES mod rbs.dna" type="DNA" seqLen="7854" ID="25" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7152-7168" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7560-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7125" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="7125..251" colors="blue-black"/><ColorRange range="6144..7124" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="7125..251" colors="blue-black"/><ColorRange range="6144..7124" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="7125" val2="7126"/><Node name="pKR13 pEX GroES mod rbs.dna" type="DNA" seqLen="7856" ID="24" circular="1" 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name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7562-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7127" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="7127..252" colors="blue-black"/><ColorRange range="6143..7124" colors="black-red"/><ColorRange range="7125..7126" colors="white"/></TopStrand><BottomStrand><ColorRange range="7127..252" colors="blue-black"/><ColorRange range="6143..7124" colors="black-red"/><ColorRange range="7125..7126" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="7127..252" colors="blue-black"/><ColorRange range="6145..7126" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="7127..252" colors="blue-black"/><ColorRange range="6145..7126" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="7127" val2="7127"/><Node name="pKR13 pEX GroES mod rbs.dna" type="DNA" seqLen="7857" ID="23" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7155-7171" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7563-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7128" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="7128..252" colors="blue-black"/><ColorRange range="6145..7126" colors="black-red"/><ColorRange range="7127..7127" colors="white"/></TopStrand><BottomStrand><ColorRange range="7128..252" colors="blue-black"/><ColorRange range="6145..7126" colors="black-red"/><ColorRange range="7127..7127" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6615..6620" colors="red"/></TopStrand><BottomStrand><ColorRange range="6615..6620" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6615"/><Node name="pKR13 pEX GroES mod rbs.dna" type="DNA" seqLen="7851" ID="22" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7149-7165" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7557-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7122" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6615..7595" colors="blue-black"/><ColorRange range="5634..6614" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6615..7595" colors="blue-black"/><ColorRange range="5634..6614" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6615" val2="6620"/><Node name="pKR13 pEX GroES mod rbs.dna" type="DNA" seqLen="7857" ID="21" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7155-7171" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7563-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7128" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="6621..7602" colors="blue-black"/><ColorRange range="5633..6614" colors="black-red"/><ColorRange range="6615..6620" colors="white"/></TopStrand><BottomStrand><ColorRange range="6621..7602" colors="blue-black"/><ColorRange range="5633..6614" colors="black-red"/><ColorRange range="6615..6620" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6615..6620" colors="red"/></TopStrand><BottomStrand><ColorRange range="6615..6620" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6615"/><Node name="pKR13 pEX GroES mod rbs.dna" type="DNA" seqLen="7851" ID="20" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7149-7165" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7557-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7122" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6615..7595" colors="blue-black"/><ColorRange range="5634..6614" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6615..7595" colors="blue-black"/><ColorRange range="5634..6614" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6615" val2="6620"/><Node name="pKR13 pEX GroES mod rbs.dna" type="DNA" seqLen="7857" ID="19" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7155-7171" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7563-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7128" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="6621..7602" colors="blue-black"/><ColorRange range="5633..6614" colors="black-red"/><ColorRange range="6615..6620" colors="white"/></TopStrand><BottomStrand><ColorRange range="6621..7602" colors="blue-black"/><ColorRange range="5633..6614" colors="black-red"/><ColorRange range="6615..6620" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..7127" colors="red"/></TopStrand><BottomStrand><ColorRange range="6107..7127" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6107"/><Node name="pKR13 pEX GroES mod rbs.dna" type="DNA" seqLen="6836" ID="18" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6134-6150" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6542-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..124" colors="blue-black"/><ColorRange range="5253..6106" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6107..124" colors="blue-black"/><ColorRange range="5253..6106" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6107" val2="7259"/><Node name="pKR13 pEX GroES mod rbs.dna" type="DNA" seqLen="7989" ID="17" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7287-7303" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7695-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6659-7260" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="7260..268" colors="blue-black"/><ColorRange range="5109..6106" colors="black-red"/><ColorRange range="6107..7259" colors="white"/></TopStrand><BottomStrand><ColorRange range="7260..268" colors="blue-black"/><ColorRange range="5109..6106" colors="black-red"/><ColorRange range="6107..7259" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="7260..268" colors="blue-black"/><ColorRange range="6262..7259" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="7260..268" colors="blue-black"/><ColorRange range="6262..7259" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="7260" val2="7260"/><Node name="pKR12 pEX∆rpsU3.dna" type="DNA" seqLen="7990" ID="16" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6659-7261" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7696-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7288-7304" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="7261..268" colors="blue-black"/><ColorRange range="6262..7259" colors="black-red"/><ColorRange range="7260..7260" colors="white"/></TopStrand><BottomStrand><ColorRange range="7261..268" colors="blue-black"/><ColorRange range="6262..7259" colors="black-red"/><ColorRange range="7260..7260" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6667..6667" colors="red"/></TopStrand><BottomStrand><ColorRange range="6667..6667" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6667"/><Node name="pKR12 pEX∆rpsU3.dna" type="DNA" seqLen="7989" ID="15" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6659-7260" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7695-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. 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transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7287-7303" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6667..6674" colors="red"/></TopStrand><BottomStrand><ColorRange range="6667..6674" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6667"/><Node name="pKR12 pEX∆rpsU3.dna" type="DNA" seqLen="7981" ID="14" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6659-7252" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7687-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7279-7295" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6667..7663" colors="blue-black"/><ColorRange range="5670..6666" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6667..7663" colors="blue-black"/><ColorRange range="5670..6666" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6667" val2="6669"/><Node name="pKR12 pEX∆rpsU3.dna" type="DNA" seqLen="7984" ID="13" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6659-7255" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7690-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7282-7298" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="6670..7667" colors="blue-black"/><ColorRange range="5669..6666" colors="black-red"/><ColorRange range="6667..6669" colors="white"/></TopStrand><BottomStrand><ColorRange range="6670..7667" colors="blue-black"/><ColorRange range="5669..6666" colors="black-red"/><ColorRange range="6667..6669" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6670..7667" colors="blue-black"/><ColorRange range="5672..6669" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6670..7667" colors="blue-black"/><ColorRange range="5672..6669" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6670" val2="6849"/><Node name="pKR12 pEX∆rpsU3.dna" type="DNA" seqLen="8164" ID="12" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6659-7435" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7870-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7462-7478" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="6850..7869" colors="blue-black"/><ColorRange range="5650..6669" colors="black-red"/><ColorRange range="6670..6849" colors="white"/></TopStrand><BottomStrand><ColorRange range="6850..7869" colors="blue-black"/><ColorRange range="5650..6669" colors="black-red"/><ColorRange range="6670..6849" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6658..7434" colors="red"/></TopStrand><BottomStrand><ColorRange range="6658..7434" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6658"/><Node name="pKR12 pEX∆rpsU3.dna" type="DNA" seqLen="7387" ID="11" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7093-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6658" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6685-6701" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..6657" colors="red"/></TopStrand><BottomStrand><ColorRange range="6107..6657" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6107"/><Node name="pKR12 pEX∆rpsU3.dna" type="DNA" seqLen="6836" ID="10" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6542-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6134-6150" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..124" colors="blue-black"/><ColorRange range="5253..6106" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6107..124" colors="blue-black"/><ColorRange range="5253..6106" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6107" val2="7157"/><Node name="pKR12 pEX∆rpsU3.dna" type="DNA" seqLen="7887" ID="9" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7158" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7593-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" 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color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7185-7201" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="7158..255" colors="blue-black"/><ColorRange range="5122..6106" colors="black-red"/><ColorRange range="6107..7157" colors="white"/></TopStrand><BottomStrand><ColorRange range="7158..255" colors="blue-black"/><ColorRange range="5122..6106" colors="black-red"/><ColorRange range="6107..7157" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6633..6641" 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allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7584-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7149" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6633..7616" colors="blue-black"/><ColorRange range="5649..6632" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6633..7616" colors="blue-black"/><ColorRange range="5649..6632" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6633" val2="6812"/><Node name="pKR10 pEX∆rpsU2.dna" type="DNA" seqLen="8058" ID="7" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7356-7372" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7764-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-7329" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="6813..7819" colors="blue-black"/><ColorRange range="5626..6632" colors="black-red"/><ColorRange range="6633..6812" colors="white"/></TopStrand><BottomStrand><ColorRange range="6813..7819" colors="blue-black"/><ColorRange range="5626..6632" colors="black-red"/><ColorRange range="6633..6812" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..7328" colors="red"/></TopStrand><BottomStrand><ColorRange range="6107..7328" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6107"/><Node name="pKR10 pEX∆rpsU2.dna" type="DNA" seqLen="6836" ID="6" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6542-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6134-6150" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..124" colors="blue-black"/><ColorRange range="5253..6106" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6107..124" colors="blue-black"/><ColorRange range="5253..6106" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6107" val2="7016"/><Node name="pKR10 pEX∆rpsU2.dna" type="DNA" seqLen="7746" ID="5" circular="1" resurrectable="1" operation="replace" hideHistory="1"><Features><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6302-7017" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7452-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6292" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7044-7060" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="FTL_0880" directionality="1" translationMW="75.07" type="CDS" allowSegmentOverlaps="0"><Segment range="6293-6297" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="G"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="7017..238" colors="blue-black"/><ColorRange range="5139..6106" colors="black-red"/><ColorRange range="6107..7016" colors="white"/></TopStrand><BottomStrand><ColorRange range="7017..238" colors="blue-black"/><ColorRange range="5139..6106" colors="black-red"/><ColorRange range="6107..7016" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..6291" colors="red"/></TopStrand><BottomStrand><ColorRange range="6107..6291" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="replace" val1="6107" val2="6240"/><Node name="pKR1 pEX∆FTL1251_B.dna" type="DNA" seqLen="7695" ID="4" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="FTL_0880" directionality="1" translationMW="75.07" type="CDS" allowSegmentOverlaps="0"><Segment range="6242-6246" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="G"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6993-7009" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6241" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7401-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6251-6966" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="6107..6240" colors="white"/></TopStrand><BottomStrand><ColorRange range="6107..6240" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6250..6965" colors="red"/></TopStrand><BottomStrand><ColorRange range="6250..6965" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6250"/><Node name="pKR1 pEX∆FTL1251.dna" type="DNA" seqLen="6979" ID="3" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="FTL_0882" directionality="2" translateFirstCodonAsMet="1" translationMW="117.16" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6251-6255" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256240" predef="GI"/><V text="3952356" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="4.1" predef="profile"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="note"><V text="/dnas_title=apolipoprotein N-acyltransferase"/></Q><Q name="product"><V text="apolipoprotein N-acyltransferase"/></Q><Q name="protein_id"><V text="YP_513602.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="V"/></Q></Feature><Feature name="FTL_0880" directionality="1" translationMW="75.07" type="CDS" allowSegmentOverlaps="0"><Segment range="6242-6246" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="G"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6277-6293" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6241" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6685-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Product"><TopStrand><ColorRange range="6250..142" colors="blue-black"/><ColorRange range="5378..6249" colors="black-red"/></TopStrand><BottomStrand><ColorRange range="6250..142" colors="blue-black"/><ColorRange range="5378..6249" colors="black-red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="remove" val1="6250" val2="7265"/><Node name="pKR1 pEX∆FTL1251.dna" type="DNA" seqLen="7995" ID="2" circular="1" resurrectable="1" operation="insert" hideHistory="1"><Features><Feature name="FTL_0880" directionality="1" translationMW="75.07" type="CDS" allowSegmentOverlaps="0"><Segment range="6242-6246" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="G"/></Q></Feature><Feature name="FTL_0880" directionality="1" translateFirstCodonAsMet="1" translationMW="319.42" type="CDS" allowSegmentOverlaps="0"><Segment range="6256-6264" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="LGM"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7293-7309" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="Francisella tularensis subsp. holarctica" type="source" allowSegmentOverlaps="0"><Segment range="6108-6241" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature name="FTL_0882" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7081-7271" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256240" predef="GI"/><V text="3952356" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="4.1" predef="profile"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="note"><V text="/dnas_title=apolipoprotein N-acyltransferase"/></Q><Q name="product"><V text="apolipoprotein N-acyltransferase"/></Q><Q name="protein_id"><V text="YP_513602.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="VPCCY*A*WRNY*RTT*GYSCDT*ANDIFK*LSYYLDEYWYVISLAIVDI*YSDWANY*REIL"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="FTL_0881" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6560-6835" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256239" predef="GI"/><V text="3952355" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0881"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513601.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="KK*KR*LHYQHYY*YL*LVLLLVKI*IIILIF*ML*KMEKILLSLLISQIVNQR*KYLGNLVLNQ**FTMIL*FFQIPILLEIIHNTQMSQY"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7701-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature></Features><HistoryColors><StrandColors type="Input"><TopStrand><ColorRange range="7266..269" colors="blue-black"/><ColorRange range="5251..6249" colors="black-red"/><ColorRange range="6250..7265" colors="white"/></TopStrand><BottomStrand><ColorRange range="7266..269" colors="blue-black"/><ColorRange range="5251..6249" colors="black-red"/><ColorRange range="6250..7265" colors="white"/></BottomStrand></StrandColors><StrandColors type="Product"><TopStrand><ColorRange range="6107..6240" colors="red"/></TopStrand><BottomStrand><ColorRange range="6107..6240" colors="red"/></BottomStrand></StrandColors></HistoryColors><InputSummary manipulation="insertAt" val1="6107"/><Node name="pKR1 pEX∆FTL1251.dna" type="DNA" seqLen="7861" ID="1" circular="1" resurrectable="1" operation="remove" hideHistory="1"><Features><Feature name="FTL_0880" directionality="1" translationMW="75.07" type="CDS" allowSegmentOverlaps="0"><Segment range="6108-6112" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="G"/></Q></Feature><Feature name="FTL_0880" directionality="1" translateFirstCodonAsMet="1" translationMW="319.42" type="CDS" allowSegmentOverlaps="0"><Segment range="6122-6130" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256238" predef="GI"/><V text="3952354" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0880"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513600.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="LGM"/></Q></Feature><Feature name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7159-7175" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="FTL_0882" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6947-7137" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256240" predef="GI"/><V text="3952356" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="4.1" predef="profile"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="note"><V text="/dnas_title=apolipoprotein N-acyltransferase"/></Q><Q name="product"><V text="apolipoprotein N-acyltransferase"/></Q><Q name="protein_id"><V text="YP_513602.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="VPCCY*A*WRNY*RTT*GYSCDT*ANDIFK*LSYYLDEYWYVISLAIVDI*YSDWANY*REIL"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="FTL_0881" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6426-6701" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256239" predef="GI"/><V text="3952355" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0881"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513601.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="KK*KR*LHYQHYY*YL*LVLLLVKI*IIILIF*ML*KMEKILLSLLISQIVNQR*KYLGNLVLNQ**FTMIL*FFQIPILLEIIHNTQMSQY"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7567-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="KanR" directionality="1" translationMW="30979.95" type="CDS" allowSegmentOverlaps="0"><Segment range="1104-1919" color="#ccffcc" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to 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gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature name="FTL_0882" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7974-8164" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256240" predef="GI"/><V text="3952356" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="4.1" predef="profile"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q><Q name="note"><V text="/dnas_title=apolipoprotein N-acyltransferase"/></Q><Q name="product"><V text="apolipoprotein N-acyltransferase"/></Q><Q name="protein_id"><V text="YP_513602.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="VPCCY*A*WRNY*RTT*GYSCDT*ANDIFK*LSYYLDEYWYVISLAIVDI*YSDWANY*REIL"/></Q></Feature><Feature name="promoter" type="promoter" allowSegmentOverlaps="0"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature name="FTL_0881" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="7453-7728" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256239" predef="GI"/><V text="3952355" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="Uniprot" predef="similar to AA sequence"/></Q><Q name="locus_tag"><V text="FTL_0881"/></Q><Q name="note"><V text="/dnas_title=hypothetical protein"/></Q><Q name="product"><V text="hypothetical protein"/></Q><Q name="protein_id"><V text="YP_513601.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="KK*KR*LHYQHYY*YL*LVLLLVKI*IIILIF*ML*KMEKILLSLLISQIVNQR*KYLGNLVLNQ**FTMIL*FFQIPILLEIIHNTQMSQY"/></Q></Feature><Feature name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="8594-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature name="FTL_0879" directionality="2" translateFirstCodonAsMet="1" translationMW="-1.00" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0"><Segment range="6113-6702" color="#993366" type="standard"/><Q name="EC_number"><V text="3.5.2.6"/></Q><Q name="codon_start"><V int="1"/></Q><Q name="db_xref"><V text="89256237" predef="GI"/><V text="3952353" predef="GeneID"/></Q><Q name="inference"><V bool="1" text="2004" predef="profile"/></Q><Q name="locus_tag"><V text="FTL_0879"/></Q><Q name="note"><V text="/dnas_title=beta-lactamase"/></Q><Q name="product"><V text="beta-lactamase"/></Q><Q name="protein_id"><V text="YP_513599.1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V 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text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature name="SacB" directionality="2" translationMW="52972.36" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038"><Segment range="2704-4038" color="#993366" type="standard"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to 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   <AlignableSequences trimStringency="Medium"><Sequence name="HT03_190613" ID="0" isTrace="1" sortOrder="8" trimmedRange="14..973"/><Sequence name="HT04_190613" ID="1" isTrace="1" sortOrder="9" trimmedRange="24..956"/></AlignableSequences>  e      BC N sre```pp23*+/*IMbqqQjZjQj^r*C< C}6         BC 햱$5{H) f/ ?[.',@@=hd4%u.wFϯ?]ϗ}7,ղF?/}~wa6ne~)Y{45j{|0;פDM53&ZgScb6afILUkF;&'.
S's=ӦTY×4(%F*,j90:uiqr	i&V0S.4S#vf1U[月
((^2 gA9DnU4rVJĪn+qB)lSHG,eyd\	Kg&?i1)Z(d>Δfb!xk9Klq4Ad߿+sE#.4B0űOA<f:of7/o7/YW=J^Qio_\KT^_yۼ7M\vcAbm`i 3 ;pcBh4;J=@'nxyn$xT@s^y$X{83gձX|炣YySpkka/%pэ@
#5reƱXi%f>3%Ƃ'UKdI%)ّ%	)*7Ɣ[Z;foeL        BC             !<AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>FivePrimePhosphorylated</UpstreamModification><DownstreamModification>FivePrimePhosphorylated</DownstreamModification></AdditionalSequenceProperties>
  <?xml version="1.0"?><Features nextValidID="18"><Feature recentID="17" name="probably start site" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6708-6710" color="#a6acb3" type="standard"/></Feature><Feature recentID="16" name="Francisella tularensis subsp. holarctica" type="source" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6108-7327" color="#000000" type="standard"/><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="15" name="FTL_1504" directionality="1" type="gene" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="6747-7327" color="#00e600" type="standard"/></Feature><Feature recentID="14" name="peroxidase/catalase" directionality="1" translationMW="21996.53" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="7349" maxFusedRunOn="7349"><Segment range="6747-7327" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MLLASSNAIAEDTKTKNDNLSPQSVDLSPLRNLNKLDSPMDKDYNYHQAFKKLDTEQLKKDMQDLLTQSQDWWPADFGNYGPFFIRLSWHDAGTYRIYDGRGGANRGQQRFSPLNSWPDNVNLDKARQLLWPIKQKYGDAVSWSDLIVLAGTVSLESMGMKPIGFAFGREDDWQGDDTNWGLSPEEIMSSNVR"/></Q></Feature><Feature recentID="13" name="M13 rev" directionality="2" type="primer_bind" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7354-7370" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="12" name="M13 fwd" directionality="1" type="primer_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6052-6068" color="#a020f0" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;common sequencing primer, one of multiple similar variants&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="11" name="rrnB T2 terminator" type="terminator" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="5497-5524" color="#ffffff" type="standard"/><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T2 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="10" name="rrnB T1 terminator" type="terminator" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="5616-5702" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Escherichia coli rrnB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;transcription terminator T1 from the &lt;i&gt;E. coli rrnB&lt;/i&gt; gene&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="9" name="AmpR promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="2296-2400" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;bla&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="8" name="oriT" type="oriT" swappedSegmentNumbering="1" allowSegmentOverlaps="0" readingFrame="-1" consecutiveTranslationNumbering="1"><Segment range="4945-5054" color="#a6acb3" type="standard"/><Q name="note"><V text="incP origin of transfer"/></Q></Feature><Feature recentID="7" name="sacB promoter" directionality="2" type="promoter" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="4126-4571" color="#ffffff" type="standard"/><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacR&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;sacB&lt;/i&gt; promoter and control region&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="6" name="KanR" directionality="1" translationMW="30979.26" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="1104-1919" color="#ccffcc" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;aph(3')-Ia&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;confers resistance to kanamycin in bacteria or G418 (Geneticin®) in eukaryotes&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;aminoglycoside phosphotransferase&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF*"/></Q></Feature><Feature recentID="5" name="SacB" directionality="2" translationMW="52971.10" type="CDS" swappedSegmentNumbering="1" allowSegmentOverlaps="0" cleavageArrows="4038" readingFrame="-1" consecutiveTranslationNumbering="1" hitsStopCodon="1"><Segment range="2704-4038" color="#993366" type="standard" translated="1"/><Segment name="signal peptide" range="4039-4125" color="#993366" type="standard" translated="1"/><Q name="codon_start"><V int="1"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;Bacillus subtilis sacB&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;negative selection marker&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="product"><V text="&lt;html&gt;&lt;body&gt;secreted levansucrase that renders bacterial growth sensitive to sucrose&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="protein_id"><V text="&lt;html&gt;&lt;body&gt;&lt;br&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="transl_table"><V int="1"/></Q><Q name="translation"><V text="MNIKKFAKQATVLTFTTALLAGGATQAFA,KETNQKPYKETYGISHITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYHIVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEWSGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDGKTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKAYYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDEIERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLKMDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSVVKDSILEQGQLTVNK*"/></Q></Feature><Feature recentID="4" name="promoter" type="promoter" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="866-1103" color="#ffffff" type="standard"/><Q name="note"><V text="groES promotoer"/></Q></Feature><Feature recentID="3" name="ColE origin" directionality="2" type="rep_origin" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="7762-288" color="#ffff00" type="standard"/><Q name="direction"><V text="LEFT"/></Q><Q name="note"><V text="ColE origin"/></Q></Feature><Feature recentID="2" name="FTL_0882" directionality="2" type="gene" swappedSegmentNumbering="1" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="7328-7332" color="#a6acb3" type="standard"/><Q name="db_xref"><V text="3952356" predef="GeneID"/></Q><Q name="locus_tag"><V text="FTL_0882"/></Q></Feature><Feature recentID="1" name="source" type="source" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" visible="0"><Segment range="1-8056" color="#ffffff" type="standard"/><Q name="note"><V text="/dnas_title=pKL80 pEX.DFTL_0880"/></Q><Q name="organism"><V text="synthetic DNA construct"/></Q></Feature><Feature recentID="0" name="mod rbs" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="6697-6702" color="#a6acb3" type="standard"/></Feature></Features>
  ;t<?xml version="1.0"?><Primers nextValidID="22"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40"/><Primer recentID="0" name="KROL203 pKR35_FR1_R" sequence="tagcatggtaaGACGAGcgctataaagttgagtttgaatacaag" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-06-05T16:23:27Z"><BindingSite location="6669-6712" boundStrand="1" annealedBases="tagcatggtaaGACGAGcgctataaagttgagtttgaatacaag" meltingTemperature="65"><Component hybridizedRange="6669-6712" bases="tagcatggtaaGACGAGcgctataaagttgagtttgaatacaag"/></BindingSite><BindingSite simplified="1" location="6669-6712" boundStrand="1" annealedBases="tagcatggtaaGACGAGcgctataaagttgagtttgaatacaag" meltingTemperature="65"><Component hybridizedRange="6669-6712" bases="tagcatggtaaGACGAGcgctataaagttgagtttgaatacaag"/></BindingSite></Primer><Primer recentID="1" name="KROL202 pKR35_FR2_F" sequence="ctttatagcgCTCGTCttaccatgctaaagaaaattataactgc" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-06-05T16:22:03Z"><BindingSite location="6686-6729" boundStrand="0" annealedBases="ctttatagcgCTCGTCttaccatgctaaagaaaattataactgc" meltingTemperature="64"><Component hybridizedRange="6686-6729" bases="ctttatagcgCTCGTCttaccatgctaaagaaaattataactgc"/></BindingSite><BindingSite simplified="1" location="6686-6729" boundStrand="0" annealedBases="ctttatagcgCTCGTCttaccatgctaaagaaaattataactgc" meltingTemperature="64"><Component hybridizedRange="6686-6729" bases="ctttatagcgCTCGTCttaccatgctaaagaaaattataactgc"/></BindingSite></Primer><Primer recentID="2" name="KROL193 cat_mod_rbs_ckF" sequence="ataattgccaaaatcagcaggccac" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-22T18:19:02Z"><BindingSite location="6961-6985" boundStrand="1" annealedBases="ataattgccaaaatcagcaggccac" meltingTemperature="61"><Component hybridizedRange="6961-6985" bases="ataattgccaaaatcagcaggccac"/></BindingSite><BindingSite simplified="1" location="6961-6985" boundStrand="1" annealedBases="ataattgccaaaatcagcaggccac" meltingTemperature="61"><Component hybridizedRange="6961-6985" bases="ataattgccaaaatcagcaggccac"/></BindingSite></Primer><Primer recentID="3" name="KROL191 pKR35_seq_2" sequence="tttatggccaatcaaacaaaaatatggt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-22T18:11:09Z"><BindingSite location="1847-1858" boundStrand="0" annealedBases="caaaaatatggt" meltingTemperature="35"><Component bases="tttatggccaatcaaa"/><Component hybridizedRange="1847-1858" bases="caaaaatatggt"/></BindingSite><BindingSite location="7132-7159" boundStrand="0" annealedBases="tttatggccaatcaaacaaaaatatggt" meltingTemperature="57"><Component hybridizedRange="7132-7159" bases="tttatggccaatcaaacaaaaatatggt"/></BindingSite><BindingSite simplified="1" location="1847-1858" boundStrand="0" annealedBases="caaaaatatggt" meltingTemperature="35"><Component bases="tttatggccaatcaaa"/><Component hybridizedRange="1847-1858" bases="caaaaatatggt"/></BindingSite><BindingSite simplified="1" location="7132-7159" boundStrand="0" annealedBases="tttatggccaatcaaacaaaaatatggt" meltingTemperature="57"><Component hybridizedRange="7132-7159" bases="tttatggccaatcaaacaaaaatatggt"/></BindingSite></Primer><Primer recentID="4" name="KROL190 pKR35_seq_1" sequence="aatttctgtttaatatataatcacttataag" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-22T18:07:12Z"><BindingSite location="6582-6612" boundStrand="0" annealedBases="aatttctgtttaatatataatcacttataag" meltingTemperature="50"><Component hybridizedRange="6582-6612" bases="aatttctgtttaatatataatcacttataag"/></BindingSite><BindingSite simplified="1" location="6582-6612" boundStrand="0" annealedBases="aatttctgtttaatatataatcacttataag" meltingTemperature="50"><Component hybridizedRange="6582-6612" bases="aatttctgtttaatatataatcacttataag"/></BindingSite></Primer><Primer recentID="5" name="KROL189 pKR35_FR2_R" sequence="atatGGTACCtctcttacattactagacattatctctt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-22T18:05:06Z"><BindingSite location="7299-7332" boundStrand="1" annealedBases="GGTACCtctcttacattactagacattatctctt" meltingTemperature="58"><Component bases="atat"/><Component hybridizedRange="7299-7332" bases="GGTACCtctcttacattactagacattatctctt"/></BindingSite><BindingSite simplified="1" location="7299-7332" boundStrand="1" annealedBases="GGTACCtctcttacattactagacattatctctt" meltingTemperature="58"><Component bases="atat"/><Component hybridizedRange="7299-7332" bases="GGTACCtctcttacattactagacattatctctt"/></BindingSite></Primer><Primer recentID="6" name="KROL188 pKR35_FR2_F" sequence="atatCTCGTCttaccatgctaaagaaaattataactgc" description="&lt;html&gt;&lt;body&gt;&#10;&#10;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-22T18:01:38Z"><BindingSite location="6696-6729" boundStrand="0" annealedBases="CTCGTCttaccatgctaaagaaaattataactgc" meltingTemperature="60"><Component bases="atat"/><Component hybridizedRange="6696-6729" bases="CTCGTCttaccatgctaaagaaaattataactgc"/></BindingSite><BindingSite simplified="1" location="6696-6729" boundStrand="0" annealedBases="CTCGTCttaccatgctaaagaaaattataactgc" meltingTemperature="60"><Component bases="atat"/><Component hybridizedRange="6696-6729" bases="CTCGTCttaccatgctaaagaaaattataactgc"/></BindingSite></Primer><Primer recentID="7" name="KROL187 pKR35_FR1_R" sequence="atatGACGAGcgctataaagttgagtttgaatacaag" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-22T17:59:17Z"><BindingSite location="6669-6701" boundStrand="1" annealedBases="GACGAGcgctataaagttgagtttgaatacaag" meltingTemperature="61"><Component bases="atat"/><Component hybridizedRange="6669-6701" bases="GACGAGcgctataaagttgagtttgaatacaag"/></BindingSite><BindingSite simplified="1" location="6669-6701" boundStrand="1" annealedBases="GACGAGcgctataaagttgagtttgaatacaag" meltingTemperature="61"><Component bases="atat"/><Component hybridizedRange="6669-6701" bases="GACGAGcgctataaagttgagtttgaatacaag"/></BindingSite></Primer><Primer recentID="8" name="KROL186 pKR35_FR1_F" sequence="atatGGATCCtagaagcgtttagctgctttagcatt" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;" dateAdded="2019-05-22T17:57:05Z"><BindingSite location="6101-6132" boundStrand="0" annealedBases="GGATCCtagaagcgtttagctgctttagcatt" meltingTemperature="63"><Component bases="atat"/><Component hybridizedRange="6101-6132" bases="GGATCCtagaagcgtttagctgctttagcatt"/></BindingSite><BindingSite simplified="1" location="6101-6132" boundStrand="0" annealedBases="GGATCCtagaagcgtttagctgctttagcatt" meltingTemperature="63"><Component bases="atat"/><Component hybridizedRange="6101-6132" bases="GGATCCtagaagcgtttagctgctttagcatt"/></BindingSite></Primer><Primer recentID="9" name="P171 RT-FTL1504-1F" sequence="AACTCATGGTGCAGTTCCAGAGGA" description="RT PCR Primers; F. tularensis LVS"><BindingSite location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite><BindingSite simplified="1" location="1794-1807" boundStrand="1" annealedBases="GCAGTTCCAAGGA" meltingTemperature="40"><Component bases="AACTCATGGT"/><Component hybridizedRange="1799-1807" bases="GCAGTTCCA"/><Component bases="G"/><Component hybridizedRange="1794-1797" bases="AGGA"/></BindingSite></Primer><Primer recentID="10" name="P508 KanCass_F" sequence="atatGATATCCAATTGtatacccttcaagctttgaaaaataaac" description="&quot;Contains EcoRV site, MfeI site&quot;"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatGATATCCAATTG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="11" name="P050 groESpromoR" sequence="atatGGATCCatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatGGATCCa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="12" name="P052 groESpromoF" sequence="atatCTGCAGatatacccttcaagctttgaaaaataaac" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F PstI; F. tularensis LVS"><BindingSite location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="55"><Component bases="atatCTGCAGa"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="13" name="P506 KanCass_F" sequence="atatCTCGAGtatacccttcaagctttgaaaaataaac" description="Contains XhoI site"><BindingSite location="853-892" boundStrand="0" annealedBases="CTCGGtatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atat"/><Component hybridizedRange="853-856" bases="CTCG"/><Component bases="A"/><Component hybridizedRange="858-859" bases="Gt"/><Component hybridizedRange="866-892" bases="atacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="865-892" boundStrand="0" annealedBases="tatacccttcaagctttgaaaaataaac" meltingTemperature="53"><Component bases="atatCTCGAG"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="14" name="P711 KanCass_F" sequence="atatCATATGtatacccttcaagctttgaaaaataaac" description="Contains NdeI site"><BindingSite location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite><BindingSite simplified="1" location="862-892" boundStrand="0" annealedBases="ATtatacccttcaagctttgaaaaataaac" meltingTemperature="54"><Component bases="atatCAT"/><Component hybridizedRange="862-863" bases="AT"/><Component bases="G"/><Component hybridizedRange="865-892" bases="tatacccttcaagctttgaaaaataaac"/></BindingSite></Primer><Primer recentID="15" name="P572 Tn_Mar3" sequence="attaagtttatttttcaaagcttgaagg" description="0"><BindingSite location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite><BindingSite simplified="1" location="870-897" boundStrand="1" annealedBases="attaagtttatttttcaaagcttgaagg" meltingTemperature="54"><Component hybridizedRange="870-897" bases="attaagtttatttttcaaagcttgaagg"/></BindingSite></Primer><Primer recentID="16" name="P457 S_pSD26_1" sequence="ttaagtttatttttcaaagcttgaaggg" description="0"><BindingSite location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite><BindingSite simplified="1" location="869-896" boundStrand="1" annealedBases="ttaagtttatttttcaaagcttgaaggg" meltingTemperature="56"><Component hybridizedRange="869-896" bases="ttaagtttatttttcaaagcttgaaggg"/></BindingSite></Primer><Primer recentID="17" name="P051 groESpromoR" sequence="atatGGATCCgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter R BamHI; F. tularensis LVS"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatGGATCCgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="18" name="P049 groESpromoF" sequence="atatCTGCAGgaatgttcataacaatcttactcctttg" description="To amplify groES (FTL_1715) promoter for outward-facing promoter F; F. tularensis LVS; Discarded! Wrong enzyme site"><BindingSite location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite><BindingSite simplified="1" location="1085-1106" boundStrand="1" annealedBases="tcataacaatcttactcctttg" meltingTemperature="51"><Component bases="atatCTGCAGgaatgt"/><Component hybridizedRange="1085-1106" bases="tcataacaatcttactcctttg"/></BindingSite></Primer><Primer recentID="19" name="P570 Tn_Mar1" sequence="tttgtcaaacaagctctcgattttaac" description="0"><BindingSite location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite><BindingSite simplified="1" location="935-961" boundStrand="1" annealedBases="tttgtcaaacaagctctcgattttaac" meltingTemperature="57"><Component hybridizedRange="935-961" bases="tttgtcaaacaagctctcgattttaac"/></BindingSite></Primer><Primer recentID="20" name="KROL7 Seq_pEX_2" sequence="TGTGGAATTGTGAGCGGATA" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="7381-7400" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="7381-7400" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite><BindingSite simplified="1" location="7381-7400" boundStrand="1" annealedBases="TGTGGAATTGTGAGCGGATA" meltingTemperature="55"><Component hybridizedRange="7381-7400" bases="TGTGGAATTGTGAGCGGATA"/></BindingSite></Primer><Primer recentID="21" name="KROL6 Seq_pEX_1" sequence="GTTTTCCCAGTCACGACGTT" description="&lt;html&gt;&lt;body&gt;&lt;/body&gt;&lt;/html&gt;"><BindingSite location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite><BindingSite simplified="1" location="6031-6050" boundStrand="0" annealedBases="GTTTTCCCAGTCACGACGTT" meltingTemperature="57"><Component hybridizedRange="6031-6050" bases="GTTTTCCCAGTCACGACGTT"/></BindingSite></Primer></Primers>
  C<Notes>
<UUID>265140f3-4473-461e-a6f2-0808c24fa2b0</UUID>
<Type>Synthetic</Type>
<ConfirmedExperimentally>0</ConfirmedExperimentally>
<Created UTC="23:34:38">2018.2.27</Created>
<LastModified UTC="14:56:52">2019.6.14</LastModified>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y    K         Unsaved Enzyme Set                                                                                                    HR  d              E                                                                                                                                                                                           <?xml version="1.0"?><CustomEnzymeSets><CustomEnzymeSet type="2" name="Unsaved Enzyme Set" enzymeNames="AfeI BamHI KpnI"/></CustomEnzymeSets>
