LOCUS       sequence-11466-e        9490 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9490)
  AUTHORS   Steen EJ, Kang Y, Bokinsky G, Hu Z, Schirmer A, McClure A, Del 
            Cardayre SB, Keasling JD.
  TITLE     Microbial production of fatty-acid-derived fuels and chemicals from 
            plant biomass.
  JOURNAL   Nature. 2010 Jan 28;463(7280):559-62.
  PUBMED    20111002
REFERENCE   2  (bases 1 to 9490)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nature."; 
            date: "2010-01-28"; volume: "463(7280)"; pages: "559-62"
COMMENT     Sequence Label: pKS17
FEATURES             Location/Qualifiers
     source          1..9490
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2455
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     primer_bind     3147..3169
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3161..3178
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3188..3207
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7551..7567)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7552..7568)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(7597..7617)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(7597..7615)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7636..7652)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7636..7652)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7649..7671)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7660..7676
                     /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)."
     promoter        complement(join(7684..7690,7691..7708,7709..7714))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7729..7750
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(7944..9134)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     8763..8782
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(9182..9187,9188..9204,9205..9210))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(9237..9255)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgggccgt ctcttgggct tgatcggcct tcttgcgcat ctcacgcgct cctgcggcgg
       61 cctgtagggc aggctcatac ccctgccgaa ccgcttttgt cagccggtcg gccacggctt
      121 ccggcgtctc aacgcgcttt gagattccca gcttttcggc caatccctgc ggtgcatagg
      181 cgcgtggctc gaccgcttgc gggctgatgg tgacgtggcc cactggtggc cgctccaggg
      241 cctcgtagaa cgcctgaatg cgcgtgtgac gtgccttgct gccctcgatg ccccgttgca
      301 gccctagatc ggccacagcg gccgcaaacg tggtctggtc gcgggtcatc tgcgctttgt
      361 tgccgatgaa ctccttggcc gacagcctgc cgtcctgcgt cagcggcacc acgaacgcgg
      421 tcatgtgcgg gctggtttcg tcacggtgga tgctggccgt cacgatgcga tccgccccgt
      481 acttgtccgc cagccacttg tgcgccttct cgaagaacgc cgcctgctgt tcttggctgg
      541 ccgacttcca ccattccggg ctggccgtca tgacgtactc gaccgccaac acagcgtcct
      601 tgcgccgctt ctctggcagc aactcgcgca gtcggcccat cgcttcatcg gtgctgctgg
      661 ccgcccagtg ctcgttctct ggcgtcctgc tggcgtcagc gttgggcgtc tcgcgctcgc
      721 ggtaggcgtg cttgagactg gccgccacgt tgcccatttt cgccagcttc ttgcatcgca
      781 tgatcgcgta tgccgccatg cctgcccctc ccttttggtg tccaaccggc tcgacggggg
      841 cagcgcaagg cggtgcctcc ggcgggccac tcaatgcttg agtatactca ctagactttg
      901 cttcgcaaag tcgtgaccgc ctacggcggc tgcggcgccc tacgggcttg ctctccgggc
      961 ttcgccctgc gcggtcgctg cgctcccttg ccagcccgtg gatatgtgga cgatggccgc
     1021 gagcggccac cggctggctc gcttcgctcg gcccgtggac aaccctgctg gacaagctga
     1081 tggacaggct gcgcctgccc acgagcttga ccacagggat tgcccaccgg ctacccagcc
     1141 ttcgaccaca tacccaccgg ctccaactgc gcggcctgcg gccttgcccc atcaattttt
     1201 ttaattttct ctggggaaaa gcctccggcc tgcggcctgc gcgcttcgct tgccggttgg
     1261 acaccaagtg gaaggcgggt caaggctcgc gcagcgaccg cgcagcggct tggccttgac
     1321 gcgcctggaa cgacccaagc ctatgcgagt gggggcagtc gaaggcgaag cccgcccgcc
     1381 tgccccccga gcctcacggc ggcgagtgcg ggggttccaa gggggcagcg ccaccttggg
     1441 caaggccgaa ggccgcgcag tcgatcaaca agccccggag gggccacttt ttgccggagg
     1501 gggagccgcg ccgaaggcgt gggggaaccc cgcaggggtg cccttctttg ggcaccaaag
     1561 aactagatat agggcgaaat gcgaaagact taaaaatcaa caacttaaaa aaggggggta
     1621 cgcaacagct cattgcggca ccccccgcaa tagctcattg cgtaggttaa agaaaatctg
     1681 taattgactg ccacttttac gcaacgcata attgttgtcg cgctgccgaa aagttgcagc
     1741 tgattgcgca tggtgccgca accgtgcggc accctaccgc atggagataa gcatggccac
     1801 gcagtccaga gaaatcggca ttcaagccaa gaacaagccc ggtcactggg tgcaaacgga
     1861 acgcaaagcg catgaggcgt gggccgggct tattgcgagg aaacccacgg cggcaatgct
     1921 gctgcatcac ctcgtggcgc agatgggcca ccagaacgcc gtggtggtca gccagaagac
     1981 actttccaag ctcatcggac gttctttgcg gacggtccaa tacgcagtca aggacttggt
     2041 ggccgagcgc tggatctccg tcgtgaagct caacggcccc ggcaccgtgt cggcctacgt
     2101 ggtcaatgac cgcgtggcgt ggggccagcc ccgcgaccag ttgcgcctgt cggtgttcag
     2161 tgccgccgtg gtggttgatc acgacgacca ggacgaatcg ctgttggggc atggcgacct
     2221 gcgccgcatc ccgaccctgt atccgggcga gcagcaacta ccgaccggcc ccggcgagga
     2281 gccgcccagc cagcccggca ttccgggcat ggaaccagac ctgccagcct tgaccgaaac
     2341 ggaggaatgg gaacggcgcg ggcagcagcg cctgccgatg cccgatgagc cgtgttttct
     2401 ggacgatggc gagccgttgg agccgccgac acgggtcacg ctgccgcgcc ggtagcactt
     2461 gggttgcgca gcaacccgta agtgcgctgt tccagactat cggctgtagc cgcctcgccg
     2521 ccctatacct tgtctgcctc cccgcgttgc gtcgcggtgc atggagccgg gccacctcga
     2581 cctgaatgga agccggcggc acctcgctaa cggattcacc gtttttatca ggctctggga
     2641 ggcagaataa atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg
     2701 gcctcaggca tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa
     2761 ccagcaatag acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa
     2821 tttgctttcg aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag
     2881 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc
     2941 agtcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     3001 aaatattaac gcttacaatt tccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3061 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3121 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3181 gcgcgcgtaa tacgactcac tatagggcga attggagcat atgttaattg gctgttttaa
     3241 tatcttcctg ctttgcaatt acgccttcaa atagttgaat ttcttcttct aaatgtgtca
     3301 gtaaattctg cattcttggc aatgcattac ggcatgcaat caaaccaact tcaagtttat
     3361 ctaaataact ggtcattgta atattcaatg cttgaccgtc taatacaatt gaagctgggt
     3421 agagtgcatc aagtttggca ccattccagt aaagtggctc tcttgggcca ggcacattgg
     3481 aaataaccag attgaaggct tggcgttttg gcatcatgcc agaaattatg ttgagtcctg
     3541 cagggccata tacgacagca ctataattta gaatctgatc gctggtcata cgtttgaagc
     3601 gttgctttga gttttgaaca ctacggcgga taatttcaag acgttgtaaa ggatcatctt
     3661 tgtgggttgc caaatttgcc agaatcatcg taatacggtt gctgacatct gaatcgtcat
     3721 tgcgaataga ggctggaacc atggcaatta atggttttga aggcaaacta ttatgactca
     3781 tcaaatacgc acgtaatgca ccagaacata ccgctagtac aacatcatta atggtcacat
     3841 tcaacgattt ggcaatatta cgaaaacgat ctaggtcaaa agactgtgct gcaaaacgtc
     3901 gcgatgagct cacacgctga ttcaaaatag aacaaggcgc ctgaaagctt gaaacatgat
     3961 caggattacg tccaatatct ttaaatactg tctgagaaag ctcttgaatg actgtgggtg
     4021 tcgcctgaag ctgactctta ataccagaca tgattttctt aattttacct gttttaggtt
     4081 ctcttaagcg ctttgcacgt tttccctcaa cacaccaagg tggcacgata cttttttctg
     4141 ttacatcatg ggagagtgat ttttcaatta accgcatacc agcaacgcca tcgaccatcg
     4201 catggtgaat tttgaagtac atggcaaaac gattgccttc aattccttca ataatattgc
     4261 aggtccacaa gggctttgcc cgatctagca gcgtactgtg ctcttgtgaa atataaataa
     4321 gcaattcacg aatacgacca ggatgaggca gtgcaatatg acgaaaatga tgatctaaat
     4381 caaactcttc atcttcatcc caaaaaagcc cattcagttt attgttgaat ggtggaacag
     4441 ggattgattt tgatatccgg atatcattca ccagatcttg aataaaggtg tctggggcgt
     4501 tatcaggaat ctgaaacaaa aataaaccac ctacatgcat aggctgttgt cttttttcta
     4561 gtgacaggaa tataaaatca atcggatgta atgggcgcat aatagtcctc ctactagttt
     4621 agaaagcgct caggaagagt tcttcaactt ctttctgatc accctgacgc gggttggtca
     4681 gagcacaagc atctttcaga gcgtggtcag caagaagcgg cacatcttct ttcttagcac
     4741 ccagctcggt caggtttgct ggaataccaa tggaagcagc cagatcgcga acagcctgaa
     4801 tggtggcttc tgcgccttct ttatcaccga gattggcgat atcgagaccc atagcaacac
     4861 caacgtcttt cagacgacca gcaacgacag aggcgttata agccagaaca tgcggaagca
     4921 gaacagcgtt gcagacacca tgcggcaggt tgtagtaacc gcccaactgg tgagccatag
     4981 catggacata accaagcgaa gcgttgttga aggccatacc agcgaggaat tgggcataag
     5041 ccatagcttc acgagccggc atatccttac cgttgtcgca agcggtcttc agattcttag
     5101 cgatcatgga agctgctttc aaagcgcaag catcggtgat cggagtagct gccgttgaag
     5161 aataagcttc aaatgcgtgg gtcagagcat ccataccggt ggcggcggtc aggccttttg
     5221 gcataccaac catcaacaga ggatcgttga cggaaaccat cggggtaacg tgacggtcaa
     5281 caatggccat cttaacgtga cggacttcat cagtgatgat gcagaaacgc gtcatttcag
     5341 aagccgtacc agccgtcgtg ttgattgaca tcaaaggcag ggcaggtttc ttagatttgt
     5401 cgataccttc gtagtctttg acttcaccac cattggttgc gaccagagcg atggctttgg
     5461 cgcagtcatg gggagaacca ccaccgaggg agatgacgaa gtctgaattg ttatccttca
     5521 ggatcttaag gccttccaga actgcggtaa cagtcgggtt cggcataacg ccatcataaa
     5581 cagcagaatt aataccctgt gctttcaaca ggtcagcaac ctgcttcaca acaccggatt
     5641 tgttcatgaa agcatcagaa acgatcagcg catttttaaa gccgctgccg ttaagatcct
     5701 tgattgcttt ttcaagcgaa ccttcgccca tttcgttgac gaaaggaata taaaaagttg
     5761 aagaagccat tactcattcc tcctccgcgg tggcggccgc tcctagagga gcttgttaac
     5821 aggcttacgg ctgttggcgg cagcaacgcg cttaccccat ttgaccaatt cttcagtgca
     5881 gtcttcacga ccgatgaagc attcgatcag ggttgggccg tcggtgtttg ccagagcaac
     5941 cttgatagct tctgccagtt cgccaccggt tttagccttc aggcctttac cagcaccgct
     6001 gtcataacca ccgttaccgt tgaacacttc catcagaccg gcataatccc agttcttgat
     6061 gttgttgtac ggaccatcat ggatcataac ttcgatggtg taaccatagt tattgatcaa
     6121 gaagatgata accggcagtt tcaggcgaac catctgagcg acttcctgag ccgtcagctg
     6181 gaaggaacca tcaccaacca tgaggatgtt gcgacgttcc ggagcaccga cggcataacc
     6241 gaaggcggca ggaacggacc aaccaatgtg accccactgc atttcatatt caacgcgagc
     6301 accgttcggg agcttcatgc gctgagcatt gaaccaagag tcaccggttt cagcaataac
     6361 cgtcgtgttc ggggtcagaa gagcttcgac ctgacgggcg atttctgcgt tgaccaacgg
     6421 agcactcgga tcagccggag cggctttctt cagttcacct gcattgaggg atttgaagaa
     6481 gtccaaagca ccggttttct tggaaacttt ctgagccaaa cgggtcagat agtctttcag
     6541 atggacgctg gggaagcgaa tgccgttaac gacgacagaa cgcggttcag cgagaaccag
     6601 tttcttagga tcaggaatat ccgtccaacc agtggtggag tagtcgttaa agacaggagc
     6661 cagagcgata accgcatcgg cttctttcat cgtcttttca acgcccggat agctgacttc
     6721 accccatgag gtaccgatgt aatgcgggtt ttcttctggg aagaagcttt ttgcagcagc
     6781 catggtagca actgcgccac caagagcatc agcaaatttg acagcagctt cttcagcacc
     6841 agctgcgcgc agcttgctgc cgacgaggac ggcaactttg tcgcggtngg cgatgaattt
     6901 cagggtttct tcaaccgctg cattcaaaga agcttcgtcg ctggcttcgt cattgaacaa
     6961 tgcgcttgcc ggtccaggag cggcgcaggg catggaagca atgttgcaag cgatttcgag
     7021 ataaaccggc ttcttctcac gaagagcagt tttaatcacg tgatcgattt tagccggagc
     7081 ttcttccggg gtataaatcg cttcagcggc ggccgtgatg ttcttggcca tttccaactg
     7141 atagtgatag tcggttttgc caagagcgtg atgcaacacg tgaccagcag cgtggtcatt
     7201 gttgttcgga gcaccggaga tcaggataac cggaaggttt tctgcatagg cgccaccgat
     7261 agcatcaaat gcggaaagcg caccgacgct gtaggtaacg acggctgctg ctgcgccttt
     7321 ggcacgagca taaccttctg cactgaaacc gcagttcagt tcgttacagc aataaacctg
     7381 ctccatgttt ttgttcaaaa gcaggttgtc aagaaggacg aggttgtagt cgcccgcgac
     7441 tgcgaagtga tgcttgagac caatctggac aagccgctcc gctaaatagg taccgacagt
     7501 ataactcatg gttaattcct cctactgcag gaattcgata tcaagcttat cgataccgtc
     7561 gacctcgagg gggggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg
     7621 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7681 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7741 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7801 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgcatttg
     7861 cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg tgaatccgtt
     7921 agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc accgcgacgc
     7981 aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa cccgttccat
     8041 gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccagtgat cgaagttagg
     8101 ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc tacctgcctg
     8161 gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag aatcataatg
     8221 gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag cgcgtcggcc
     8281 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     8341 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     8401 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     8461 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     8521 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg ctctccctta
     8581 tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
     8641 gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     8701 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     8761 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     8821 acgatgcgtc cggcgtagag gatccacagg acgggtgtgg tcgccatgat cgcgtagtcg
     8881 atagtggctc caagtagcga agcgagcagg actgggcggc ggccaaagcg gtcggacagt
     8941 gctccgagaa cgggtgcgca tagaaattgc atcaacgcat atagcgctag cagcacgcca
     9001 tagtgactgg cgatgctgtc ggaatggacg atatcccgca agaggcccgg cagtaccggc
     9061 ataaccaagc ctatgcctac agcatccagg gtgacggtgc cgaggatgac gatgagcgca
     9121 ttgttagatt tcatacacgg tgcctgactg cgttagcaat ttaactgtga taaactaccg
     9181 cattaaagct tatcgatgat aagctgtcaa acatgagaat tcttgaagac gaaagggcct
     9241 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     9301 tggcactttt cggggaaatg tgcgcgcccg cgttcctgct ggcgctgggc ctgtttctgg
     9361 cgctggactt cccgctgttc cgtcagcagc ttttcgccca cggccttgat gatcgcggcg
     9421 gccttggcct gcatatcccg attcaacggc cccagggcgt ccagaacggg cttcaggcgc
     9481 tcccgaaggt
//
