LOCUS       Exported                8217 bp ds-DNA     circular SYN 06-JUN-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8217)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 21, 2019 from SnapGene 4.1.9
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8217
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=pKL126 pEXDFTL_0402"
     source          6108..6854
                     /label=Francisella tularensis subsp. holarctica
     source          6864..7488
                     /label=Francisella tularensis subsp. holarctica
     primer_bind     4..26
                     /label=P025 pEX18KanSeq1F
                     /note="P025 pEX18KanSeq1F"
     primer_bind     474..493
                     /label=P037 pEX18KanSeq9F
                     /note="P037 pEX18KanSeq9F"
     primer_bind     733..755
                     /label=P026 pEX18KanSeq2F
                     /note="P026 pEX18KanSeq2F"
     promoter        866..1103
                     /note="groES promotoer"
     CDS             1104..1919
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1583..1607)
                     /label=P027 pEX18KanSeq3F
                     /note="P027 pEX18KanSeq3F"
     promoter        complement(2296..2400)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             complement(2704..4125)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     primer_bind     2968..2994
                     /label=P029 pEX18KanSeq5F
                     /note="P029 pEX18KanSeq5F"
     primer_bind     3649..3675
                     /label=P030 pEX18KanSeq6F
                     /note="P030 pEX18KanSeq6F"
     promoter        complement(4126..4571)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     primer_bind     4390..4416
                     /label=P031 pEX18KanSeq7F
                     /note="P031 pEX18KanSeq7F"
     oriT            4945..5054
                     /note="incP origin of transfer"
     primer_bind     5113..5137
                     /label=P033 pEX18KanSeq8F
                     /note="P033 pEX18KanSeq8F"
     terminator      5497..5524
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      5616..5702
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     6013..6036
                     /label=P023 pEX18SeqF2
                     /note="P023 pEX18SeqF2"
     primer_bind     6037..6056
                     /label=P021 pEX18SeqF1
                     /note="P021 pEX18SeqF1"
     primer_bind     6052..6068
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6102..6135
                     /label=P789 DFTL_1548FR1F
     misc_feature    6108..6854
                     /label=DFTL_1548_FR1
     CDS             complement(6108..6664)
                     /codon_start=1
                     /label=DNA gyrase subunit B
                     /translation="MSENKAYDSSSIKVLKGLDAVRKRPGMYIGDTDDGSGLHHMVFEV
                     VDNAIDEALAGYCDDIKVIINKDGSVSVSDNGRGIPTDIHKEEGRSAAEVILTVLHAGG
                     KFDDNSYKVSGGLHGVGVSVVNALSEQLKLTIYRNGIEHYQEYVHGVPQFPLKEVGSTD
                     KTGTLITFKPSKETFSFVDFDY"
     gene            complement(6108..6664)
                     /label=FTL_1547
     primer_bind     6516..6542
                     /label=P793 DFTL_1548_int1
     primer_bind     complement(6826..6862)
                     /label=P790 DFTL_1548FR1R
     CDS             6846..6854
                     /codon_start=1
                     /label=FTL_1545 hypothetical protein
                     /translation="LKK"
     primer_bind     6855..6893
                     /label=P791 DFTL_1548FR2F
     gene            6855..6859
                     /locus_tag="FTL_0402"
                     /label=FTL_0402
                     /db_xref="GeneID:3952329"
     misc_feature    6864..7488
                     /label=DFTL_1548_FR2
     CDS             6864..6872
                     /codon_start=1
                     /label=hypothetical protein
                     /translation="PL"
     CDS             6906..7488
                     /codon_start=1
                     /label=tRNA-dihydrouridine synthase A
                     /translation="MKITTRKICIAPMLDWTDRHYRYMMRLITRHTMLYTEMVTLNAIL
                     HGNKEFLLKYNPQENPVTLQLGGCVPKDFISCGKLAEELDYNEININVGCPSERVKKGN
                     FGLSLMAEPELVADCVKAMKDNLNIPISVKCRIGYDHNDSYEELYNFVDKQVQAGIDYL
                     CIHARKGWLSGLSPKENRTIPELKYDVVYSI"
     gene            6906..7488
                     /label=FTL_1549
     primer_bind     complement(7461..7494)
                     /label=P792 DFTL_1548FR2R
     gene            complement(7489..7493)
                     /locus_tag="FTL_0404"
                     /label=FTL_0404
                     /db_xref="GeneID:3952331"
     primer_bind     complement(7515..7531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7526..7551)
                     /label=P024 pEX18SeqR2
                     /note="P024 pEX18SeqR2"
     protein_bind    7539..7555
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     complement(7548..7567)
                     /label=P022 pEX18SeqR1
                     /note="P022 pEX18SeqR1"
     promoter        complement(7563..7593)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7608..7629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(join(7917..8217,1..288))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
       61 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
      121 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
      181 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      241 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
      301 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
      361 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
      421 tcaatctaaa gtatatatga gtaaacttgg tctgacaatc gatcaagggt tggtttgcgc
      481 attcacagtt ctccgcaaga attgattggc tccaattctt ggagtggtga atccgttagc
      541 gaggtgccgc cggcttccat tcaggtcgag gtggcccggc tccatgcacc gcgacgcaac
      601 gcggggaggc agacaaggta tagggcggcg cctacaatcc atgccaaccc gttccatgtg
      661 ctcgccgagg cggcataaat cgccgtgacg atcagcggtc caatgatcga agttaggctg
      721 gtaagagccg cgagcgatcc ttgaagctgt ccctgatggt cgtcatctac ctgcctggac
      781 agcatggcct gcaacgcggg catcccgatg ccgccggaag cgagaagaat cataatgggg
      841 aaggccatcc agcctcgcgt cgatctatac ccttcaagct ttgaaaaata aacttaatta
      901 ttatatatgt tatttagcta gtttttttaa ttaaagttaa aatcgagagc ttgtttgaca
      961 aaaaaacaaa aaaatttctt gaaaattttt tttttgactc aatatctaga cttgcaagag
     1021 cttggaactt tgagattgtt ctaagatgca tacaaattca aaatgcttaa acaaaaataa
     1081 tttaacaaag gagtaagatt gttatgagcc atattcaacg ggaaacgtct tgctcgaggc
     1141 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     1201 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt
     1261 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     1321 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     1381 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     1441 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     1501 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     1561 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     1621 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     1681 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     1741 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     1801 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     1861 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     1921 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     1981 gctttgttga ataaatcgaa cttttgctga gttgaaggat cagatcacgc atcttcccga
     2041 caacgcagac cgttccgtgg caaagcaaaa gttcaaaatc accaactggt ccacctacaa
     2101 caaagctctc atcaacgata tcccgcaaga ggcccggcag taccggcata accaagccta
     2161 tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg ttagatttca
     2221 tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat taaagctaat
     2281 cggttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     2341 tctcatgagc ggatacatat ttgaatgtat ttagagaaat aaacaaatag gggttccgcg
     2401 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     2461 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2521 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2581 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2641 ctatgcggca tcagagcaga ttgtactgag agtgcaccat aatcggcatt ttcttttgcg
     2701 tttttatttg ttaactgtta attgtccttg ttcaaggatg ctgtctttga caacagatgt
     2761 tttcttgcct ttgatgttca gcaggaagct aggcgcaaac gttgattgtt tgtctgcgta
     2821 gaatcctctg tttgtcatat agcttgtaat cacgacattg tttcctttcg cttgaggtac
     2881 agcgaagtgt gagtaagtaa aggttacatc gttaggatca agatccattt ttaacacaag
     2941 gccagttttg ttcagcggct tgtatgggcc agttaaagaa ttagaaacat aaccaagcat
     3001 gtaaatatcg ttagacgtaa tgccgtcaat cgtcattttt gatccgcggg agtcagtgaa
     3061 cagataccat ttgccgttca ttttaaagac gttcgcgcgt tcaatttcat ctgttactgt
     3121 gttagatgca atcagcggtt tcatcacttt tttcagtgtg taatcatcgt ttagctcaat
     3181 cataccgaga gcgccgtttg ctaactcagc cgtgcgtttt ttatcgcttt gcagaagttt
     3241 ttgactttct tgacggaaga atgatgtgct tttgccatag tatgctttgt taaataaaga
     3301 ttcttcgcct tggtagccat cttcagttcc agtgtttgct tcaaatacta agtatttgtg
     3361 gcctttatct tctacgtagt gaggatctct cagcgtatgg ttgtcgcctg agctgtagtt
     3421 gccttcatcg atgaactgct gtacattttg atacgttttt ccgtcaccgt caaagattga
     3481 tttataatcc tctacaccgt tgatgttcaa agagctgtct gatgctgata cgttaacttg
     3541 tgcagttgtc agtgtttgtt tgccgtaatg tttaccggag aaatcagtgt agaataaacg
     3601 gatttttccg tcagatgtaa atgtggctga acctgaccat tcttgtgttt ggtcttttag
     3661 gatagaatca tttgcatcga atttgtcgct gtctttaaag acgcggccag cgtttttcca
     3721 gctgtcaata gaagtttcgc cgactttttg atagaacatg taaatcgatg tgtcatccgc
     3781 atttttagga tctccggcta atgcaaagac gatgtggtag ccgtgatagt ttgcgacagt
     3841 gccgtcagcg ttttgtaatg gccagctgtc ccaaacgtcc aggccttttg cagaagagat
     3901 atttttaatt gtggacgaat cgaactcagg aacttgatat ttttcatttt tttgctgttc
     3961 agggatttgc agcatatcat ggcgtgtaat atgggaaatg ccgtatgttt ccttatatgg
     4021 cttttggttc gtttctttcg caaacgcttg agttgcgcct cctgccagca gtgcggtagt
     4081 aaaggttaat actgttgctt gttttgcaaa ctttttgatg ttcatcgttc atgtctcctt
     4141 ttttatgtac tgtgttagcg gtctgcttct tccagccctc ctgtttgaag atggcaagtt
     4201 agttacgcac aataaaaaaa gacctaaaat atgtaagggg tgacgccaaa gtatacactt
     4261 tgccctttac acattttagg tcttgcctgc tttatcagta acaaacccgc gcgatttact
     4321 tttcgacctc attctattag actctcgttt ggattgcaac tggtctattt tcctcttttg
     4381 tttgatagaa aatcataaaa ggatttgcag actacgggcc taaagaacta aaaaatctat
     4441 ctgtttcttt tcattctctg tattttttat agtttctgtt gcatgggcat aaagttgcct
     4501 ttttaatcac aattcagaaa atatcataat atctcatttc actaaataat agtgaacggc
     4561 aggtatatgt gatgggttaa aaaggatcga tcctctagct agagtcgatc ttcgccagca
     4621 gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg gtcgtcggtg agccagagtt
     4681 tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg ggccagctcg cggacgtgct
     4741 catagtccac gacgcccgtg attttgtagc cctggccgac ggccagcagg taggccgaca
     4801 ggctcatgcc ggctgccgcc gccttttcct caatcgctct tcgttcgtct ggaaggcagt
     4861 acaccttgat aggtgggctg cccttcctgg ttggcttggt ttcatcagcc atccgcttgc
     4921 cctcatctgt tacgccggcg gtagccggcc agcctcgcag agcaggattc ccgttgagca
     4981 ccgccaggtg cgaataaggg acagtgaaga aggaacaccc gctcgcgggt gggcctactt
     5041 cacctatcct gcccggctga cgccgttgga tacaccaagg aaagtctaca cgaacccttt
     5101 ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac cgaaaaaatc gctataatga
     5161 ccccgaagca gggttatgca gcggaaaagc gctgcttccc tgctgttttg tggaatatct
     5221 accgactgga aacaggcaaa tgcaggaaat tactgaactg aggggacagg cgagagacga
     5281 tgccaaagag ctacaccgac gagctggccg agtgggttga atcccgcgcg gccaagaagc
     5341 gccggcgtga tgaggctgcg gttgcgttcc tggcggtgag ggcggatgtc gatatgcggt
     5401 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgcata tttgaatgta
     5461 tttagaaaaa taaacaaaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     5521 ttctgcttaa tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     5581 cgttgcttcg caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     5641 accgacaaac aacagataaa acgaaaggcc cagtctttcg actgagcctt tcgttttatt
     5701 tgatgcctgg cagttcccta ctctcgcatg gggagacccc acactaccat cggcgctacg
     5761 gcgtttcact tctgagttcg gcatggggtc aggtgggacc accgcgctac tgccgccagg
     5821 caaattctgt tttatcagac cgcttctgcg ttctgattta atctgtatca ggctgaaaat
     5881 cttctctcat ccgccaaaac agccaagctc gccattcgcc attcaggctg cgcaactgtt
     5941 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     6001 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     6061 cggccagtgc caagcttgca tgcctgcagg tcgactctag aggatcctca taatcaaagt
     6121 caacaaatga aaacgtttct ttactcggtt taaaagtaat tagtgttccg gtcttatctg
     6181 ttgaaccaac ttcttttaat ggaaactgtg gtactccatg aacatactct tgatagtgct
     6241 cgataccgtt tctataaatt gttaacttta attgttctga aagcgcattt acaactgata
     6301 caccaacacc atgtaaaccg cctgatactt tataagagtt atcatcaaac ttaccacctg
     6361 catgcaatac tgtcaaaata acctctgctg cagagcgccc ttcttcttta tgaatatctg
     6421 taggaatacc tctaccatta tctgatacag atacagagcc atccttattt ataataactt
     6481 taatatcatc acaataacca gctaaggctt catcgatagc attatcaaca acctcaaata
     6541 ccatatgatg taagccgcta ccatcatcgg tatcaccaat atacattcca ggtctttttc
     6601 taacagcatc taaaccttta agtaccttaa tactcgatga gtcataagct ttattctcag
     6661 acattaacaa actccaaatt tatcgttata ttttaaaatt caatcattaa gtattataac
     6721 acaaataaat cttgattatt aatgttgata acaatgtgag ttaataaaca ttgagggttt
     6781 ttcattttat ccaaaaatgc taatattaac tctaacagtt ttgcagatag gttcattttg
     6841 aaattttgaa aaaggcggcc gctcctttat aattttataa caacttaaat ctcgtagctt
     6901 ttaaaatgaa aataacaact agaaaaattt gtatagcacc aatgcttgac tggactgata
     6961 ggcactatcg ttacatgatg cgtctaatca ccaggcatac aatgttatat actgaaatgg
     7021 ttactttaaa cgcaatctta catggtaata aagaattttt gcttaaatat aatcctcaag
     7081 aaaatcctgt gacactacag ttaggtggtt gcgttccaaa agactttatc agttgtggca
     7141 agcttgccga agaacttgac tataatgaaa ttaacataaa tgttggctgc ccaagtgaaa
     7201 gagttaaaaa aggtaatttt ggcttatcat taatggcaga gccagagctt gtagcagact
     7261 gtgtcaaagc tatgaaagat aacctaaata tcccaatttc agttaaatgt cgtattggct
     7321 atgatcataa tgatagctat gaagaactat ataattttgt tgataagcaa gttcaagcag
     7381 gaatagatta cctttgtatt catgcacgta aaggttggct tagcggtcta tcaccaaaag
     7441 aaaatcgtac catacctgag cttaaatatg atgttgtcta cagtatcagg taccgagctc
     7501 gaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     7561 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     7621 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     7681 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     7741 gcttcctcgc tcactgaccc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     7801 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     7861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     7921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     7981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     8041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     8101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     8161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaac
//
