LOCUS       sequence-11467-e        6749 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 6749)
  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 6749)
  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: pKS18
FEATURES             Location/Qualifiers
     source          1..6749
                     /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(4810..4826)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4811..4827)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(4856..4876)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(4856..4874)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4895..4911)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4895..4911)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4908..4930)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4919..4935
                     /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(4943..4949,4950..4967,4968..4973))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4988..5009
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(5203..6393)
                     /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     6022..6041
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        complement(join(6441..6446,6447..6463,6464..6469))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(6496..6514)
                     /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 attggagctc ttagtagcgc atggtggagg
     3241 aggcgcggaa gtaagacagg aacttgtagc acagagaaac aacgaagtac caaatattgc
     3301 gggccatctg agagcgggca atgaaaatcc gccaaatcag aataactaaa atggtgttaa
     3361 agccataccg aatattacgc agcttgttca gatgcttacg tgccgccggc aggccgctca
     3421 tttcttcatt cagcacgtac ttcttggtgc ccatgcaata gttttcaata tactccgccc
     3481 aatgtaactg ccgcacatcg atgttaaagg ttttcttatc ctccggattc agttggttca
     3541 tcagcatgtt cacgttatcg gtattccaca cccaggaatt agaggtaaaa tattccagaa
     3601 aaaccattgc tttatgcagg cgggtgatag ttttcatcat ccgcggactg cggccagtca
     3661 tgcgtaaata aatatcatat aagaatgccg gcgctttgtg ggacacagcg atccagtaat
     3721 gatacagcag gtgattactt gttaaattta cgtttggacg acgaaatgct tgttccagtg
     3781 gattacgctt aaaagttgaa ataacgtgat attcgacttc gccccagtga aatggattcg
     3841 tgctcccggt ggtacaattg taaaccatga tgttacgcgg acggtttacc ccgctatacc
     3901 aggccgcagc cagtgaagta ttaacgacga catccaccgg taccagatcg gctaaagcat
     3961 tgttggaggc gcgcatagta cgcagaatgc ctttacccgc agcgatgaac agaccactcg
     4021 ggccgttaaa gttatcgatc caacccggaa acggctcttt ccagctagca cctacaatag
     4081 acgggcgcac gatagcaacg ttcagtttcg caccttcttg ctgaactaca tattctgcta
     4141 aggctttcgt ataaatatat gtattcgggc gatcaccgat cagtttaggc gtaatgtcat
     4201 tcaccagccc gtcatccatc cactccagac tgtcgatcag ttttttaggg tccacaggtg
     4261 gaggatatac cacctcatcg atatgtttac ggttacagta tgcgtaagcg gttgaaacat
     4321 gcatgaacac ttccaggttt ttcatttgct gcgctaacag aattaactgg cgcgtcgcaa
     4381 tgacattcag ttggacggca tcccgcaggt tttcgttgaa ccgaactgtc gctgcacagt
     4441 gaaagattac atttgttgag tcgatgatga tttctttatc ttcttcgctc agtgccagct
     4501 taggctgggt cagttcgcta ttaatcgcaa taattttctc acggaaatcc gggttttcgt
     4561 cgcgcagacg gtcaaacagt ttagaactta aaatttcctc aacgcgttcc tgcggcgtct
     4621 ggcccgcttt ctgacgcacc agcacgtaga ctgaattgac acgtgggcag ctacgcagta
     4681 atttttccag caggactttg cccagaaacc ctgtggcacc cgttaacagg atgtttttac
     4741 cctcataata ctctgggatg ctcaccatgg ttaattcctc ctactgcagg aattcgatat
     4801 caagcttatc gataccgtcg acctcgaggg ggggcccggt acccagcttt tgttcccttt
     4861 agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     4921 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     4981 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5041 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5101 tgcgtattgg gcgcatttgc gcattcacag ttctccgcaa gaattgattg gctccaattc
     5161 ttggagtggt gaatccgtta gcgaggtgcc gccggcttcc attcaggtcg aggtggcccg
     5221 gctccatgca ccgcgacgca acgcggggag gcagacaagg tatagggcgg cgcctacaat
     5281 ccatgccaac ccgttccatg tgctcgccga ggcggcataa atcgccgtga cgatcagcgg
     5341 tccagtgatc gaagttaggc tggtaagagc cgcgagcgat ccttgaagct gtccctgatg
     5401 gtcgtcatct acctgcctgg acagcatggc ctgcaacgcg ggcatcccga tgccgccgga
     5461 agcgagaaga atcataatgg ggaaggccat ccagcctcgc gtcgcgaacg ccagcaagac
     5521 gtagcccagc gcgtcggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     5581 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     5641 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     5701 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     5761 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     5821 cggtcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta gtaggttgag
     5881 gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc
     5941 cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga gcccgaagtg
     6001 gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt
     6061 ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atccacagga cgggtgtggt
     6121 cgccatgatc gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg
     6181 gccaaagcgg tcggacagtg ctccgagaac gggtgcgcat agaaattgca tcaacgcata
     6241 tagcgctagc agcacgccat agtgactggc gatgctgtcg gaatggacga tatcccgcaa
     6301 gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc
     6361 gaggatgacg atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt
     6421 taactgtgat aaactaccgc attaaagctt atcgatgata agctgtcaaa catgagaatt
     6481 cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     6541 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcgcccgc gttcctgctg
     6601 gcgctgggcc tgtttctggc gctggacttc ccgctgttcc gtcagcagct tttcgcccac
     6661 ggccttgatg atcgcggcgg ccttggcctg catatcccga ttcaacggcc ccagggcgtc
     6721 cagaacgggc ttcaggcgct cccgaaggt
//
