LOCUS       Exported                8189 bp DNA     circular SYN 30-SEP-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pJB866 vector (V013302)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8189)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(159..239)
                     /label=Pm promoter
                     /note="The bacterial Pm promoter is activated by XylS in 
                     the presence of benzoate or m-toluate (Marques et al., 
                     1999)."
     CDS             960..1925
                     /codon_start=1
                     /gene="xylS"
                     /product="XylS regulator encoded by the Pseudomonas putida 
                     TOL plasmid pWWO"
                     /label=XylS
                     /note="stimulates transcription from the Pm promoter in the
                     presence of aromatic compounds such as m-toluate or 
                     benzoate"
                     /translation="MDFCLLNEKSQIFVHAEPYAVSDYVNQYVGTHSIRLPKGGRPAGR
                     LHHRIFGCLDLCRISYGGSVRVISPGLETCYHLQIILKGHCLWRGHGQEHYFAPGELLL
                     LNPDDQADLTYSEDCEKFIVKLPSVVLDRACSDNNWHKPREGIRFAARHNLQQLDGFIN
                     LLGLVCDEAEHTKSMPRVQEHYAGIIASKLLEMLGSNVSREIFSKGNPSFERVVQFIEE
                     NLKRNISLERLAELAMMSPRSLYNLFEKHAGTTPKNYIRNRKLESIRACLNDPSANVRS
                     ITEIALDYGFLHLGRFAENYRSAFGELPSDTLRQCKKEVA"
     primer_bind     2362..2378
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     oriT            2466..2574
                     /note="incP origin of transfer"
     CDS             complement(3037..3687)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     CDS             3793..4992
                     /codon_start=1
                     /gene="tetA"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKPNIPLIVILSTVALDAVGIGLIMPVLPGLLRDLVHSNDVTAHY
                     GILLALYALVQFACAPVLGALSDRFGRRPILLVSLAGATVDYAIMATAPFLWVLYIGRI
                     VAGITGATGAVAGAYIADITDGDERARHFGFMSACFGFGMVAGPVLGGLMGGFSPHAPF
                     FAAAALNGLNFLTGCFLLPESHKGERRPLRREALNPLASFRWARGMTVVAALMAVFFIM
                     QLVGQVPAALWVIFGEDRFHWDATTIGISLAAFGILHSLAQAMITGPVAARLGERRALM
                     LGMIADGTGYILLAFATRGWMAFPIMVLLASGGIGMPALQAMLSRQVDEERQGQLQGSL
                     AALTSLTSIVGPLLFTAIYAASITTWNGWAWIAGAALYLLCLPALRRGLWSGAGQRADR
                     "
     rep_origin      5577..6108
                     /label=oriV
                     /note="incP origin of replication"
     CDS             6501..7649
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
ORIGIN
        1 cgcgcgcacc gtacgtctcg aggaattcct gcaggatatc tggatccacg aagcttccca
       61 tggtgacgtc accggttcta gatacctagg tgagctctgg taccggcggc cgcgcggccg
      121 caattcacat gttcatgact ccattattat tgtttctgtt gcataaagcc taaggggtag
      181 gcctttctag agatagccat tttttgcact cctgtatccg cttcttgcaa ggctggactt
      241 atccctatca aaccggacac ccggtcgttg gtcagatcaa gctaattccc atgcatggtg
      301 aaaacggggg cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc
      361 acccagggat tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc
      421 aggttttcac cgtaacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg
      481 tcgtggtatt cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa
      541 caagggtgaa cactatccca tatcaccagc tcaccgtctt tcattgccat acgtaattcc
      601 ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta
      661 tttttcttta cggtctttaa aaaggccgta atatccagca gatccccttt atccgccaat
      721 tcgtcgccgc atgccccgac agcacgaact tctggtgttc tcgcttctta aaaagaacgt
      781 cttcgttctg cttggcgtta tttttgcttg gaaaagtggt cactgattgc aaaaaggatg
      841 gcgcaacgtg gcaatggggg taacccgtat acgcatcacg tcgagatgca ttttcatcga
      901 cttggcgcct ttctacatca caccaagcag cccacattaa aataagagaa ccgtgaacta
      961 tggatttttg cttattgaac gagaaaagtc agatcttcgt ccacgccgag ccctatgcag
     1021 tctccgatta tgttaaccag tatgtcggta cgcactctat tcgcctgccc aagggcgggc
     1081 gcccggcagg caggctgcac cacagaatct tcggatgcct cgacctgtgt cgaatcagct
     1141 acggcggtag cgtgagggta atctcgcctg gattagagac ctgttatcat ctgcaaataa
     1201 tactcaaagg ccattgcctg tggcgtggcc atggccagga gcactatttt gcgccgggcg
     1261 aactattgct gctcaatccg gatgaccaag ccgacctgac ctattcagaa gattgcgaga
     1321 aatttatcgt taaattgccc tcagtggtcc ttgatcgggc atgcagtgac aacaattggc
     1381 acaagccgag ggagggtatc cgtttcgccg cgcgacacaa tctccagcaa ctcgatggct
     1441 ttatcaatct actcgggtta gtttgtgacg aagcggaaca tacaaagtcg atgcctcggg
     1501 tccaagagca ctatgcgggg atcatcgctt ccaagctgct cgaaatgctg ggcagcaatg
     1561 tcagccgtga aattttcagc aaaggtaacc cgtctttcga gcgagtcgtt caattcattg
     1621 aggagaatct caaacggaat atcagccttg agcggttagc ggagctggcg atgatgagtc
     1681 cacgctcgct ctacaatttg ttcgagaagc atgccggcac cacgccgaag aactacatcc
     1741 gcaaccgcaa gctcgaaagc atccgcgcct gcttgaacga tcccagtgcc aatgtgcgta
     1801 gtataactga gatagcccta gactacggct tcttacattt gggacgcttc gctgaaaact
     1861 ataggagcgc gttcggcgag ttgccttccg acaccctgcg tcaatgcaaa aaggaagtgg
     1921 cttgattacg aacgtagccg aagaagggat gggttggcat cgccccggct ttcttagaca
     1981 ctctccaagc tctgaaatag cgttttacaa actccactgg ctatagtgcc ggcgtttgcc
     2041 cgcgctctcc gtggccgcgt cccaatgcag gtcgggttcc tcgacccgaa gtgcccttgg
     2101 gcatctccac ttcagcgtct gaccttgctg gccagttcgc cgccgatgcc gaagtggccc
     2161 ttggccatct ccgtgataat cactcgcacg ctggtcagcg gcgcatccag ggagcgcgag
     2221 atggcctcgc tgacttcccg aatgagggtt tccttctgct cgtcgctgcg gccttcaagg
     2281 atgtggatct gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
     2341 ggttttccca gtcacgacgt tgtaaaacga cggccagtga attaattctt gaagacgaaa
     2401 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagag
     2461 cttatcggcc agcctcgcag agcaggattc ccgttgagca ccgccaggtg cgaataaggg
     2521 acagtgaaga aggaacaccc gctcgcgggt gggcctactt cacctatcct gcccggctga
     2581 cgccgttgga tacaccaagg aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt
     2641 gcgaaaaagg atggatatac cgaaaaaatc gctataatga ccccgaagca gggttatgca
     2701 gcggaaaaga tccgtcgacc tgcagcgatc cgtcgatcta tctcatctgc gcaaggcaga
     2761 acgtgaagac ggccgccctg gacctcgccc gcgagcgcca ggcgcacgag gccggcgcgc
     2821 ggacccgcgc cacggcccac gagcggacgc cgcagcagga gcgccagaag gccgccagag
     2881 aggccgagcg cggccgtgag gcttggacgc tagggcaggg catgaaaaag cccgtagcgg
     2941 gctgctacgg gcgtctgacg cggtggaaag ggggagggga tgttgtctac atggctctgc
     3001 tgtagtgagt gggttgcgct ccggcagcgg tcctgatcaa tcgtcaccct ttctcggtcc
     3061 ttcaacgttc ctgacaacga gcctcctttt cgccaatcca tcgacaatca ccgcgagtcc
     3121 ctgctcgaac gctgcgtccg gaccggcttc gtcgaaggcg tctatcgcgg cccgcaacag
     3181 cggcgagagc ggagcctgtt caacggtgcc gccgcgctcg ccggcatcgc tgtcgccggc
     3241 ctgctcctca agcacggccc caacagtgaa gtagctgatt gtcatcagcg cattgacggc
     3301 gtccccggcc gaaaaacccg cctcgcagag gaagcgaagc tgcgcgtcgg ccgtttccat
     3361 ctgcggtgcg cccggtcgcg tgccggcatg gatgcgcgcg ccatcgcggt aggcgagcag
     3421 cgcctgcctg aagctgcggg cattcccgat cagaaatgag cgccagtcgt cgtcggctct
     3481 cggcaccgaa tgcgtatgat tctccgccag catggcttcg gccagtgcgt cgagcagcgc
     3541 ccgcttgttc ctgaagtgcc agtaaagcgc cggctgctga acccccaacc gttccgccag
     3601 tttgcgtgtc gtcagaccgt ctacgccgac ctcgttcaac aggtccaggg cggcacggat
     3661 cactgtattc ggctgcaact ttgtcatgct tgacacttta tcactgataa acataatatg
     3721 tccaccaact tatcagtgat aaagaatccg cgcgttcaat cggaccagcg gaggctggtc
     3781 cggaggccag acgtgaaacc caacataccc ctgatcgtaa ttctgagcac tgtcgcgctc
     3841 gacgctgtcg gcatcggcct gattatgccg gtgctgccgg gcctcctgcg cgatctggtt
     3901 cactcgaacg acgtcaccgc ccactatggc attctgctgg cgctgtatgc gttggtgcaa
     3961 tttgcctgcg cacctgtgct gggcgcgctg tcggatcgtt tcgggcggcg gccaatcttg
     4021 ctcgtctcgc tggccggcgc cactgtcgac tacgccatca tggcgacagc gcctttcctt
     4081 tgggttctct atatcgggcg gatcgtggcc ggcatcaccg gggcgactgg ggcggtagcc
     4141 ggcgcttata ttgccgatat cactgatggc gatgagcgcg cgcggcactt cggcttcatg
     4201 agcgcctgtt tcgggttcgg gatggtcgcg ggacctgtgc tcggtgggct gatgggcggt
     4261 ttctcccccc acgctccgtt cttcgccgcg gcagccttga acggcctcaa tttcctgacg
     4321 ggctgtttcc ttttgccgga gtcgcacaaa ggcgaacgcc ggccgttacg ccgggaggct
     4381 ctcaacccgc tcgcttcgtt ccggtgggcc cggggcatga ccgtcgtcgc cgccctgatg
     4441 gcggtcttct tcatcatgca acttgtcgga caggtgccgg ccgcgctttg ggtcattttc
     4501 ggcgaggatc gctttcactg ggacgcgacc acgatcggca tttcgcttgc cgcatttggc
     4561 attctgcatt cactcgccca ggcaatgatc accggccctg tagccgcccg gctcggcgaa
     4621 aggcgggcac tcatgctcgg aatgattgcc gacggcacag gctacatcct gcttgccttc
     4681 gcgacacggg gatggatggc gttcccgatc atggtcctgc ttgcttcggg tggcatcgga
     4741 atgccggcgc tgcaagcaat gttgtccagg caggtggatg aggaacgtca ggggcagctg
     4801 caaggctcac tggcggcgct caccagcctg acctcgatcg tcggacccct cctcttcacg
     4861 gcgatctatg cggcttctat aacaacgtgg aacgggtggg catggattgc aggcgctgcc
     4921 ctctacttgc tctgcctgcc ggcgctgcgt cgcgggcttt ggagcggcgc agggcaacga
     4981 gccgatcgct gatcgtggaa acgataggga cggatcgctg caggtcgacg gatcgcaaac
     5041 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     5101 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     5161 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     5221 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     5281 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     5341 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     5401 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     5461 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     5521 gggggatcag gaccgctgcc ggagcgcaac ccactcacta cagcagagcc atgtagggcc
     5581 gccggcgttg tggatacctc gcggaaaact tggccctcac tgacagatga ggggcggacg
     5641 ttgacacttg aggggccgac tcacccggcg cggcgttgac agatgagggg caggctcgat
     5701 ttcggccggc gacgtggagc tggccagcct cgcaaatcgg cgaaaacgcc tgattttacg
     5761 cgagtttccc acagatgatg tggacaagcc tggggataag tgccctgcgg tattgacact
     5821 tgaggggcgc gactactgac agatgagggg cgcgatcctt gacacttgag gggcagagtg
     5881 ctgacagatg aggggcgcac ctattgacat ttgaggggct gtccacaggc agaaaatcca
     5941 gcatttgcaa gggtttccgc ccgtttttcg gccaccgcta acctgtcttt taacctgctt
     6001 ttaaaccaat atttataaac cttgttttta accagggctg cgccctgtgc gcgtgaccgc
     6061 gcacgccgaa ggggggtgcc cccccttctc gaaccctccc ggcccgctaa cgcgggcctc
     6121 ccatcccccc aggggctgcg cccctcggcc gcgaacggcc tcaccccaaa aatggcagcc
     6181 acgtagaaag ccagtccgca gaaacggtgc tgaccccgga tgaatgtcag ctactgggct
     6241 atctggacaa gggaaaacgc aagcgcaaag agaaagcagg tagcttgcag tgggcttaca
     6301 tggcgatagc tagactgggc ggttttatgg acagcaagcg aaccggaatt gccagctggg
     6361 gcgccctctg gtaaggttgg gaagccctgc aaagtaaact ggatggcttt cttgccgcca
     6421 aggatctgat ggcgcagggg atcaagatcg acggatcgat ccggggaatt aattccgggg
     6481 caatcccgca aggagggtga atgaatcgga cgtttgaccg gaaggcatac aggcaagaac
     6541 tgatcgacgc ggggttttcc gccgaggatg ccgaaaccat cgcaagccgc accgtcatgc
     6601 gtgcgccccg cgaaaccttc cagtccgtcg gctcgatagt ccagcaagct acggccaaga
     6661 tcgagcgcga cagcgtgcaa ctggctcccc ctgccctgcc cgcgccatcg gccgccgtgg
     6721 agcgttcgcg tcgtctcgaa caggaggcgg caggtttggc gaagtcgatg accatcgaca
     6781 cgcgaggaac tatgacgacc aagaagcgaa aaaccgccgg cgaggacctg gcaaaacagg
     6841 tcagcgaggc caagcaggcc gcgttgctga aacacacgaa gcagcagatc aaggaaatgc
     6901 agctttcctt gttcgatatt gcgccgtggc cggacacgat gcgagcgatg ccaaacgaca
     6961 cggcccgctc tgccctgttc accacgcgca acaagaaaat cccgcgcgag gcgctgcaaa
     7021 acaaggtcat tttccacgtc aacaaggacg tgaagatcac ctacaccggc gtcgagctgc
     7081 gggccgacga tgacgaactg gtgtggcagc aggtgttgga gtacgcgaag cgcaccccta
     7141 tcggcgagcc gatcaccttc acgttctacg agctttgcca ggacctgggc tggtcgatca
     7201 atggccggta ttacacgaag gccgaggaat gcctgtcgcg cctacaggcg acggcgatgg
     7261 gcttcacgtc cgaccgcgtt gggcacctgg aatcggtgtc gctgctgcac cgcttccgcg
     7321 tcctggaccg tggcaagaaa acgtcccgtt gccaggtcct gatcgacgag gaaatcgtcg
     7381 tgctgtttgc tggcgaccac tacacgaaat tcatatggga gaagtaccgc aagctgtcgc
     7441 cgacggcccg acggatgttc gactatttca gctcgcaccg ggagccgtac ccgctcaagc
     7501 tggaaacctt ccgcctcatg tgcggatcgg attccacccg cgtgaagaag tggcgcgagc
     7561 aggtcggcga agcctgcgaa gagttgcgag gcagcggcct ggtggaacac gcctgggtca
     7621 atgatgacct ggtgcattgc aaacgctagg gccttgtggg gtcagttccg gctgggggtt
     7681 cagcagccac ctgcatcgca agctagcttg agagggtcag ctttatgctt gtaaaccgtt
     7741 ttgtgaaaaa atttttaaaa taaaaaaggg gacctctagg gtccccaatt aattagtaat
     7801 ataatctatt aaaggtcatt caaaaggtca tccaccggat caattcccct gctcgcgcag
     7861 gctgggtgcc aagctctcgg gtaacatcaa ggcccgatcc ttgcaaggta gtcggcaaat
     7921 aatgtctaac aattcgttca agccgacgcc gcttcgcggc gcggcttaac tcaagcgtta
     7981 gatgcactaa gcacataatt gctcacagcc aaactatcag gtcaagtctg cttttattat
     8041 ttttaagcgt gcataataag ccctacacaa attgggagat atatcatgaa aggctggctt
     8101 tttcttgtta tcgcaatagt tggcgaagta atcgcaacat ccgcattaaa atctagcgag
     8161 ggctttacta agctgatctt cgaatgcat
//
