LOCUS       Exported                8249 bp ds-DNA     circular SYN 24-OCT-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8249)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Apr 21, 2021 from SnapGene 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8249
                     /organism="synthetic DNA construct"
                     /note="/dnas_title=pKL80 pEX.DFTL_0880"
     source          6108..6867
                     /organism="synthetic DNA construct"
                     /label=Francisella tularensis subsp. holarctica
     source          6877..7520
                     /organism="synthetic DNA construct"
                     /label=Francisella tularensis subsp. holarctica
     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"
     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"
     promoter        complement(4126..4571)
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     oriT            4945..5054
                     /note="incP origin of transfer"
     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     6032..6051
                     /label=KROL6 Seq_pEX_1
     primer_bind     6052..6068
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6102..6134
                     /label=KROL8 DFTL0766_FR1F
     CDS             6108..6804
                     /codon_start=1
                     /label=VacJ lipoprotein
                     /translation="FQNLFEPSRVANDLFQGEWDYAGDDSMRFLANTTLGIAGFFDVAN
                     SWFNLPMRYHQDFAVTLHKWGVYKKGYASPYIVWPIFGPGTIEGISDSVDALFNPLSYL
                     FLAPGVGTFTAYVVNYSVYGLYYTNQGVSYLPAYSNLEEVSIDPYAALRNAYIQNYDYG
                     MAKVLKQKDPALDSTQQSNQAILSVLGISDKNGKSEIASSGESFVQASSDTPAVLKSSL
                     SSVNQVAEV"
     gene            6108..6804
                     /label=FTL_0765
     primer_bind     6713..6736
                     /label=KROL14 CkDpKR3intF
     primer_bind     complement(6840..6875)
                     /label=KROL9 DFTL0766_FR1R
     CDS             6859..6867
                     /codon_start=1
                     /label=gamma-glutamyltranspeptidase
                     /translation="MRR"
     gene            6859..6867
                     /label=FTL_0766
     primer_bind     6867..6907
                     /label=KROL10 DFTL0766_FR2F
     gene            6868..6872
                     /locus_tag="FTL_0880"
                     /label=FTL_0880
                     /db_xref="GeneID:3952354"
     CDS             6877..6885
                     /codon_start=1
                     /label=gamma-glutamyltranspeptidase
                     /translation="GY"
     gene            6877..6885
                     /label=FTL_0766
     CDS             complement(6901..7520)
                     /codon_start=1
                     /label=hypothetical protein
                     /translation="NGQIAIKLAEFINEIHATDISSAQLAKAFKHNSIKYFKADEANSM
                     FADRCVDLITVSQVAHWFDMSKFEKECLRILKPNGIVAIWAYHHNISVNTAVEIIYQEF
                     YRTIRPYFPQGREHIDNFYKDINIDLPKLDSPEFRQTKEMNFDDFIEYLKSFSAYAEYL
                     RNHNKCPIVKLGFYDKFRESWGDSKKVYTVVWPIIFKCYINK"
     gene            complement(6901..7520)
                     /label=FTL_0767
     primer_bind     complement(7493..7526)
                     /label=KROL11 DFTL0766_FR2R
     gene            complement(7521..7525)
                     /locus_tag="FTL_0882"
                     /label=FTL_0882
                     /db_xref="GeneID:3952356"
     primer_bind     complement(7547..7563)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7575..7594)
                     /label=KROL7 Seq_pEX_2
     rep_origin      complement(join(7955..8249,1..288))
                     /direction=LEFT
                     /label=ColE origin
                     /note="ColE origin"
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 aggatccttt tcaaaacttg
     6121 tttgagccta gtagagtagc aaatgatcta ttccaaggtg aatgggacta tgcaggtgat
     6181 gatagtatgc gttttttagc taatacgact cttggaattg caggcttctt tgatgttgca
     6241 aatagttggt ttaatctacc aatgagatat caccaagatt ttgctgtgac attacataag
     6301 tggggagtat ataaaaaagg ctatgcatca ccgtatatag tttggccaat ttttggtcct
     6361 ggaactatag aggggattag tgatagtgtt gatgcgctat ttaacccact ttcatattta
     6421 tttttagcac caggggtagg cacatttact gcttatgttg ttaactatag tgtctatgga
     6481 ctatattata caaatcaagg cgtttcatat ttaccagcat attcaaattt agaagaagtt
     6541 tctattgatc catatgctgc tttgcgtaat gcctacatac aaaattatga ttatggtatg
     6601 gcaaaagtcc ttaaacaaaa agatccagcg ttagattcaa cacagcaaag taatcaagcg
     6661 atactaagtg ttttaggaat tagtgataaa aatggtaaat cagaaatagc atcaagtggc
     6721 gaaagttttg tgcaagcctc atcagataca ccagctgtgc ttaaaagcag tctttcaagt
     6781 gtaaaccagg ttgctgaagt ttaattaaat attatgatgt aatttttaaa tattttttgg
     6841 tttatttttg tttctttaat gcgtcgtgcg gccgccggtt attaaacact ttaatagatt
     6901 ttatttgttg atataacatt taaatataat tggccaaacg acagtatata cttttttact
     6961 gtctccccat gattctctga atttatcata aaatcctagt ttgactatag gacatttatt
     7021 atgatttctg aggtattcgg cataagcaga aaaagatttg agatattcaa taaaatcatc
     7081 aaaattcatt tctttagttt gcctaaactc tggagaatcc agcttaggta aatcgatatt
     7141 aatatcttta taaaagttat caatatgttc tctaccttgt gggaagtatg ggcggatagt
     7201 tctataaaac tcctggtaaa taatttcaac cgcagtattg acgctgatat tatggtgata
     7261 tgcccaaata gcaacaattc cattaggttt aagtatgcgt aggcactctt tttcaaattt
     7321 actcatatca aaccaatgag caacctgtga aactgttatt agatcaacac atcgatcagc
     7381 aaacatacta ttagcttcat ctgctttaaa atatttaata ctgttatgtt taaaagcttt
     7441 agctagttgt gcactgctaa tatcagtagc atggatttca ttgataaatt cagctaattt
     7501 gattgctatt tgaccattac ggtaccgagc tcgaattcgt aatcatgtca tagctgtttc
     7561 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     7621 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     7681 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     7741 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac ccgctgcgct
     7801 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     7861 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     7921 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     7981 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     8041 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     8101 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     8161 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     8221 agcccgaccg ctgcgcctta tccggtaac
//
