	   SnapGene      gcgcGGTACCATCATTTTTCCCTATCAGTGATAGAGATTGAATTCTTAGATATTTTGATATATAATTAGATACTAATCCCTATCAGTGATAGAGAATTGATAACTTATAAAGAATTAAATTTTCTTTTGTATGCTAACTTGATTGCTAATATGAATTATACTAGTCCCTATCAGTGATAGAGACTAGTTAGTATGTTGATTATAATGATTAGAGTTTTAAATAATGGAGGTAACAATatgccaagcgttagaattaaagaaagagaaccttttgatgtcgctcttagaagatttaaaagatcttgcgaaaaggctggaatagtatctgaactacgtcgtagagaatactttgaaaagccaacttgggcacgcaaaagaaaaaaaactgccgctgttaaaagagctcataagagcaatataatagttaaaaggGCGGCCGCaTATACTGATATTGAAATGAATAGATTAGGTAAATAAGGATCCACTA  	                                                                                                                                                                                                                                                                                    	               	                                                                                                                                                    
                                    
                                                                                                                                                                                                                                                                                                                                                                                               l   ;              m 7    ! [  T     2_x ' ] o   <V G   0 8 D W b   #    ! *            U       Q     A            N         0        r  n 8         d         6   Z   S \ n  Uw " / C  )    T    @          c 5 m  (v    b	 l  '4au &3`tyz{|} t   ~   	    =W ( ? ^ p u    J      e  Hg 	            X  > = c j    1 9 ;    E  X $ _   @ q       3 v " )    
  Q   +C K      L p          G   I f  Zh       
   k  :       &  F^
  7 V a               /        
  . B Y    R    ?    k s   % > I     d         : < i    2   P  B K   F  Y   j % 6 `   ] ~   . - A     r   $ h      5 } \  #   4 w |    x      * y        O   R         D   + z ,         L            1       P   M   q         S       s   f      	  o      9 E      H     O   J  -i , g  { [                   e       N           M      d  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,GABNNNNNRTC,GAYNNNNNRTC,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                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                                  )k  )o  )q  )q  )  )  *?D h    ' t   l) ln) #'    ,  S  MD  S          *7c: < i       L( ? ^ p u    r   $ 
L  z      07zXZ  ִF !   t/ ] ˇf{3(1v%f M*	mj)p80*jBJA>f"AvӒp&s{3Th$h|=27|r6
ͽ;29TmgX#3b5p=~(qQE{tG:Ёa=a6'!rRg#H&=ҭy¦IwI?`5      B[&;u   ^Dg    YZ  <7zXZ  ִF !   t/] 3D].?I*ә_.ZvќP,Vj,ު~mC<ґ8hlFOlY.k(DW`!
 	KJ9rEPܤ$FcprMUq3+Xfg=Gjur#5Ɇca>48r7Nn2h݊rNgKä>֒ڟSgad2Kƾz$ 9VK;KMZcfJD*i~;G:b$]~[:XY`PyqlːQ񮫿BMd%3A-l&%@8N|y%T2@3eHIkͱ\I@-HMbZRGa]2'ߌe]1'OBq|wƝ}Y &s>[B[pʂ<Hf!EDK[Й~59ՈH9Iaz7Ln-H)^_7M`il?<q[{32ijFQI_.:Ƹ?$.!Z0y;(w<sO[%6'SgnM-"jVr1cT!~; ~#E8e{ͤ3_=L*%ehBxVֱđ5"m>UК[M;C'֞%nT"33}iOMԑb]<)n93#GQ|3-96ap+]SKN-|k6ȃaƓ_m+_K~{~3ɜ0}Yѝ:pƞ.e_$%@F2
6K_Z.+M7;F*}@^LŻkZV@cG{pX6<4n5%q `+ eE{7w"\Rp_;jfvZh1I_!@}ً=eI7A@{XZ5;#aXCbQ<7Ş=(x8'1|01`)gP:JTj[D.2\
^TYRW[ǉ[Ϸ EX⺌'ڞ49+لn|̴[cj{4%v^Ny) U)hka 
'  ffg    YZ  z      07zXZ  ִF !   t/ ] ˇf{3(1v%f M*	mj)p80*jBJA>f"AvӒp&s{3Th$h|=27|r6
ͽ;29TmgX#3b5p=~(qQE{tG:Ёa=a6'!rRg#H&=ҭy¦IwI?`5      B[&;u   ^Dg    YZ  <7zXZ  ִF !   t/] 3D].?I*ә_.ZvќP,Vj,ު~mC<ґ8hlFOlY.k(DW`!
 	KJ9rEPܤ$FcprMUq3+Xfg=Gjur#5Ɇca>48r7Nn2h݊rNgKä>֒ڟSgad2Kƾz$ 9VK;KMZcfJD*i~;G:b$]~[:XY`PyqlːQ񮫿BMd%3A-l&%@8N|y%T2@3eHIkͱ\I@-HMbZRGa]2'ߌe]1'OBq|wƝ}Y &s>[B[pʂ<Hf!EDK[Й~59ՈH9Iaz7Ln-H)^_7M`il?<q[{32ijFQI_.:Ƹ?$.!Z0y;(w<sO[%6'SgnM-"jVr1cT!~; ~#E8e{ͤ3_=L*%ehBxVֱđ5"m>UК[M;C'֞%nT"33}iOMԑb]<)n93#GQ|3-96ap+]SKN-|k6ȃaƓ_m+_K~{~3ɜ0}Yѝ:pƞ.e_$%@F2
6K_Z.+M7;F*}@^LŻkZV@cG{pX6<4n5%q `+ eE{7w"\Rp_;jfvZh1I_!@}ً=eI7A@{XZ5;#aXCbQ<7Ş=(x8'1|01`)gP:JTj[D.2\
^TYRW[ǉ[Ϸ EX⺌'ڞ49+لn|̴[cj{4%v^Ny) U)hka 
'  ffg    YZ  7zXZ  ִF !   t/] ˇf{3(1v%f M*	mj)p80*i	NY]QF7&!n/U"]PCDg'05c)e*f)OTIݥKkS$VYBv[e[`皡F48ۘM,74צd}"	TCDRI; p R+`КEޗsAFsv)_Rj@(nbl1
vH
!v.xq'P	5+jBD6Lǹ9#*LbpMN	!<~Fjo|BMu(Bpr\4w)2q&aq; #+EoQhFuG"j)"՗M0 *2]|op.qxBWnwWmڸ˼Js.е[ո
<p}8x|3]R.tG`\%Bx~u^ yJT+;F&[hZN | V q=[T|6["EDv׀jg[l	0TؕkB*T%-iTu|22;߰ @pZCG$0 Z~ּ	L$+`ӑ$6h,yр&唑TGYtXKbΣ(_><)^km|yxx.^&L'
\Ო̹FI%9s.,܇aO[LFXJ!1qgqE&>&#1xslAtBƳ.7,E ɟEnefς1q*]69 s#_%uĘRM    |AY   Tg    YZ   -<AlignableSequences trimStringency="Medium"/>  <AdditionalSequenceProperties><UpstreamStickiness>0</UpstreamStickiness><DownstreamStickiness>0</DownstreamStickiness><UpstreamModification>Unmodified</UpstreamModification><DownstreamModification>Unmodified</DownstreamModification></AdditionalSequenceProperties>
  [<?xml version="1.0"?><Features nextValidID="10"><Feature recentID="0" name="VSV-G" directionality="1" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="442-474" color="#993366" type="standard"/><Q name="note"><V text="VSV-G"/></Q></Feature><Feature recentID="1" name="rpsU2" directionality="1" type="gene" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="238-432" color="#00e600" type="standard"/></Feature><Feature recentID="2" name="30S ribosomal protein S21" directionality="1" translationMW="7840.31" type="CDS" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1" maxRunOn="477" maxFusedRunOn="477"><Segment range="238-432" color="#ff0000" type="standard" translated="1"/><Q name="codon_start"><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="transl_table"><V int="1"/></Q><Q name="translation"><V text="MPSVRIKEREPFDVALRRFKRSCEKAGIVSELRRREYFEKPTWARKRKKTAAVKRAHKSNIIVKR"/></Q></Feature><Feature recentID="3" name="tet operator" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="165-183" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;tetracycline repressor TetR&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;tetO&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;bacterial operator O&lt;sub&gt;2&lt;/sub&gt; for the &lt;i&gt;tetR&lt;/i&gt; and &lt;i&gt;tetA&lt;/i&gt; genes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="4" name="tet operator" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="77-95" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;tetracycline repressor TetR&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;tetO&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;bacterial operator O&lt;sub&gt;2&lt;/sub&gt; for the &lt;i&gt;tetR&lt;/i&gt; and &lt;i&gt;tetA&lt;/i&gt; genes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature><Feature recentID="5" name="TSS" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="76-76" color="#a6acb3" type="standard"/></Feature><Feature recentID="6" name="-10" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="61-66" color="#a6acb3" type="standard"/></Feature><Feature recentID="7" name="-35" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="38-43" color="#a6acb3" type="standard"/></Feature><Feature recentID="8" name="RBS" type="misc_feature" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="226-230" color="#a6acb3" type="standard"/></Feature><Feature recentID="9" name="tet operator" type="protein_bind" allowSegmentOverlaps="0" consecutiveTranslationNumbering="1"><Segment range="19-37" color="#31849b" type="standard"/><Q name="bound_moiety"><V text="&lt;html&gt;&lt;body&gt;tetracycline repressor TetR&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="gene"><V text="&lt;html&gt;&lt;body&gt;&lt;i&gt;tetO&lt;/i&gt;&lt;/body&gt;&lt;/html&gt;"/></Q><Q name="note"><V text="&lt;html&gt;&lt;body&gt;bacterial operator O&lt;sub&gt;2&lt;/sub&gt; for the &lt;i&gt;tetR&lt;/i&gt; and &lt;i&gt;tetA&lt;/i&gt; genes&lt;/body&gt;&lt;/html&gt;"/></Q></Feature></Features>
  E<?xml version="1.0"?><Primers nextValidID="1"><HybridizationParams minContinuousMatchLen="10" allowMismatch="1" minMeltingTemperature="40" showAdditionalFivePrimeMatches="1" minimumFivePrimeAnnealing="15"/><Primer recentID="0" name="KROL414 tetO_rpsU2_BamHI_R" sequence="atatGGATCCTTATTTACCTAATCTATTCATT" dateAdded="2021-03-01T17:48:13Z"><BindingSite location="455-483" boundStrand="1" annealedBases="tGGATCCTTATTTACCTAATCTATTCATT" meltingTemperature="55"><Component bases="ata"/><Component hybridizedRange="455-483" bases="tGGATCCTTATTTACCTAATCTATTCATT"/></BindingSite><BindingSite simplified="1" location="455-483" boundStrand="1" annealedBases="tGGATCCTTATTTACCTAATCTATTCATT" meltingTemperature="55"><Component bases="ata"/><Component hybridizedRange="455-483" bases="tGGATCCTTATTTACCTAATCTATTCATT"/></BindingSite></Primer></Primers>
  2<Notes>
<UUID>5bf6fe91-4c8f-4ef8-9c97-c294588e9816</UUID>
<Type>Natural</Type>
<Created UTC="17:44:14">2021.6.25</Created>
<LastModified UTC="17:44:20">2021.6.25</LastModified>
<CreatedBy>Kathryn Ramsey</CreatedBy>
<SequenceClass>UNA</SequenceClass>
<TransformedInto>unspecified</TransformedInto>
</Notes>
  Y    K         Unique 6+ Cutters                                                                                            d      HO                E             Y? P                                                                                                                                                              )<?xml version="1.0"?><CustomEnzymeSets/>
   3<?xml version="1.0"?><EnzymeVisibilities vals=""/>
