LOCUS       V015273                 8244 bp    DNA     circular SYN 26-NOV-2019
DEFINITION  Exported.
ACCESSION   V015273
VERSION     V015273
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8244)
  AUTHORS   Calles B, Goni-Moreno A, de Lorenzo V.
  TITLE     Digitalizing heterologous gene expression in Gram-negative bacteria
            with a portable on/off module
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8244)
  AUTHORS   Calles B, Goni-Moreno A, de Lorenzo V.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-AUG-2019) Systems Biology Program, Centro Nacional de
            Biotecnologia, Darwin, 3, Cantoblanco, Madrid 28049, Spain
REFERENCE   3  (bases 1 to 8244)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (28-AUG-2019) Systems Biology Program, Centro Nacional de
            Biotecnologia, Darwin, 3, Cantoblanco, Madrid 28049, Spain"
FEATURES             Location/Qualifiers
     source          1..8244
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(22..984)
                     /label="XylS"
                     /note="XylS regulator encoded by the Pseudomonas putida TOL
                     plasmid pWWO"
     promoter        1884..1964
                     /label="Pm promoter"
                     /note="The bacterial Pm promoter is activated by XylS in
                     the presence of benzoate or m-toluate (Marques et al.,
                     1999)."
     regulatory      1999..2005
                     /regulatory_class="ribosome_binding_site"
     CDS             2017..3096
                     /label="lacI"
                     /note="lac repressor"
     CDS             3097..3114
                     /label="6xHis"
                     /note="6xHis affinity tag"
     CDS             3140..4789
                     /gene="ceaC"
                     /label="Colicin-E3"
                     /note="Colicin-E3 from Escherichia coli. Accession#:
                     P00646"
     misc_feature    4793..4849
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     regulatory      4882..4911
                     /label="synthetic T500 terminator"
                     /note="synthetic T500 terminator"
                     /regulatory_class="terminator"
     terminator      complement(4918..4945)
                     /label="T7Te terminator"
                     /note="phage T7 early transcription terminator"
     terminator      4985..5028
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     regulatory      5046..5154
                     /note="anti LacI sRNA; MicC-based"
                     /regulatory_class="other"
     regulatory      complement(5155..5206)
                     /label="PA1/O4s"
                     /note="PA1/O4s"
                     /regulatory_class="promoter"
     terminator      5217..5311
                     /label="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
     CDS             5547..6335
                     /label="SmR"
                     /note="aminoglycoside adenylyltransferase (Murphy, 1985)"
     oriT            6484..6592
                     /label="oriT"
                     /note="incP origin of transfer"
     CDS             complement(6606..7265)
                     /label="pBBR1 Rep"
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     rep_origin      complement(7266..8037)
                     /direction=LEFT
                     /label="pBBR1 oriV"
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1
                     Rep protein for replication"
     terminator      complement(8152..8238)
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
ORIGIN
        1 ttaattaaac gttcgtaatc aagccacttc ctttttgcat tgacgcaggg tgtcggaagg
       61 caactcgccg aacgcgctcc tatagttttc agcgaagcgt cccaaatgta agaagccgta
      121 gtctagggct atctcagtta tactacgcac attggcactg ggatcgttca agcaggcgcg
      181 gatgctttcg agcttgcggt tgcggatgta gttcttcggc gtggtgccgg cgtgcttctc
      241 gaacaaattg tagagcgagc gtggactcat catcgccagc tccgctaacc gctcaaggct
      301 gatattccgt ttgagattct cctcaatgaa ttgaacgact cgctcgaaag acgggttacc
      361 tttgctgaaa atttcacggc tgacattgct gcccagcatt tcgagcagct tggaagcgat
      421 gatccccgca tagtgctctt ggacccgagg catcgacttt gtatgttccg cttcgtcaca
      481 aactaacccg agtagattga taaagccatc gagttgctgg agattgtgtc gcgcggcgaa
      541 acggataccc tccctcggct tgtgccaatt gttgtcactg cacgcccgat caaggaccac
      601 tgagggcaat ttaacgataa atttctcgca atcttctgaa taggtcaggt cggcttggtc
      661 atccggattg agcagcaata gttcgcccgg cgcaaaatag tgctcctggc catggccacg
      721 ccacaggcaa tggcctttga gtattatttg cagatgataa caggtctcta atccaggcga
      781 gattaccctc acgctaccgc cgtagctgat tcgacacaga tcgaggcatc cgaagattct
      841 gtggtgcagc ctgcctgccg ggcgcccgcc cttgggcagg cgaatagagt gcgtaccgac
      901 atactggtta acataatcgg agactgcata gggctcggcg tggacgaaga tctgactttt
      961 ctcgttcaat aagcaaaaat ccatagttca cggttctctt attttaatgt gggctgcttg
     1021 gtgtgatgta gaaaggcgcc aagtcgatga aaatgcatct cgacgtgatg cgtatacggg
     1081 ttacccccat tgccacgttg cgccatcctt tttgcaatca gtgaccactt ttccaagcaa
     1141 aaataacgcc aagcagaacg aagacgttct ttttaagaag cgagaacacc agaagttcgt
     1201 gctgtcgggg catggggcga cgaattggcg gataaagggg atctgctgga tattacggcc
     1261 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1321 gcccgcctga tgaatgctca tccgtaatta cgtatggcaa tgaaagacgg tgagctggtg
     1381 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1441 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1501 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1561 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1621 gacaacttct tcgcccccgt tttcaccatg catgggaatt agcttgatct gaccaacgac
     1681 cggtagcgga gctatccaac ggcggtatac caggaaaaca cacagcaggt acatcagaac
     1741 agtaccatga ctgaagaaca aatagttttt tcctgatcca taaagcagaa cggcctgctc
     1801 catgacaaat ctggctcccc aactaatgcc ccatgcagcc agcataacca gcataaagtg
     1861 cagtgtccgg tttgataggg ataagtccag ccttgcaaga agcggataca ggagtgcaaa
     1921 aaatggctat ctctagtaag gcctacccct taggctttat gcaacagaaa caataataat
     1981 ggagtcatga ccatgcctag gaggaaaaaa catatggtga aaccagtaac gttatacgat
     2041 gtcgcagagt atgccggtgt ctcttatcag accgtttccc gcgtggtgaa ccaggccagc
     2101 cacgtttctg cgaaaacgcg ggaaaaagtg gaagcggcga tggcggagct gaattacatt
     2161 cccaaccgcg tggcacaaca actggcgggc aaacagtcgt tgctgattgg cgttgccacc
     2221 tccagtctgg ccctgcacgc gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat
     2281 caactgggtg ccagcgtggt ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa
     2341 gcggcggtgc acaatcttct cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg
     2401 gatgaccagg atgccattgc tgtggaagct gcctgcacta atgttccggc gttatttctt
     2461 gatgtctctg accagacacc catcaacagt attattttct cccatgaaga cggtacgcga
     2521 ctgggcgtgg agcatctggt cgcattgggt caccagcaaa tcgcgctgtt agcgggccca
     2581 ttaagttctg tctcggcgcg tctgcgtctg gctggctggc ataaatatct cactcgcaat
     2641 caaattcagc cgatagcgga acgggaaggc gactggagtg ccatgtccgg ttttcaacaa
     2701 accatgcaaa tgctgaatga gggcatcgtt cccactgcga tgctggttgc caacgatcag
     2761 atggcgctgg gcgcaatgcg cgccattacc gagtccgggc tgcgcgttgg tgcggatatc
     2821 tcggtagtgg gatacgacga taccgaagac agctcatgtt atatcccgcc gttaaccacc
     2881 atcaaacagg attttcgcct gctggggcaa accagcgtgg accgcttgct gcaactctct
     2941 cagggccagg cggtgaaggg caatcagctg ttgcccgtct cactggtgaa aagaaaaacc
     3001 accctggcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     3061 ctggcacgac aggtttcccg actggaaagc gggcagcatc atcaccacca tcattaggat
     3121 ggtggtgatg ataatggcta gcggtggcga tggacgcggc cataacacgg gcgcgcatag
     3181 cacaagtggt aacattaatg gtggcccgac cgggcttggt gtaggtggtg gtgcttctga
     3241 tggctccgga tggagttcgg aaaataaccc gtggggtggt ggttccggta gcggcattca
     3301 ctggggtggt ggttccggtc atggtaatgg cggggggaat ggtaattccg gtggtggttc
     3361 gggaacaggc ggtaatctgt cagcagtagc tgcgccagtg gcatttggtt ttccggcact
     3421 ttccactcca ggagctggcg gtctggcggt cagtatttca gcgggagcat tatcggcagc
     3481 tattgctgat attatggctg ccctgaaagg accgtttaaa tttggtcttt ggggggtggc
     3541 tttatatggt gtattgccat cacaaatagc gaaagatgac cccaatatga tgtcaaagat
     3601 tgtgacgtca ttacccgcag atgatattac tgaatcacct gtcagttcat tacctctcga
     3661 taaggcaaca gtaaacgtaa atgttcgtgt tgttgatgat gtaaaagacg agcgacagaa
     3721 tatttcggtt gtttcaggtg ttccgatgag tgttccggtg gttgatgcaa aacctaccga
     3781 acgtccgggt gtttttacgg catcaattcc aggtgcacct gttctgaata tttcagttaa
     3841 taacagtacg ccagcagtac agacattaag cccaggtgtt acaaataata ctgataagga
     3901 tgttcgcccg gcaggattta ctcagggtgg taataccagg gatgcagtta ttcgattccc
     3961 gaaggacagc ggtcataatg ccgtatatgt ttcagtgagt gatgttctta gccctgacca
     4021 ggtaaaacaa cgtcaagatg aagaaaatcg ccgtcagcag gaatgggatg ctacgcatcc
     4081 ggttgaagcg gctgagcgaa attatgaacg cgcgcgtgca gagctgaatc aggcaaatga
     4141 agatgttgcc agaaatcagg agcgacaggc taaagctgtt caggtttata attcgcgtaa
     4201 aagcgaactt gatgcagcga ataaaactct tgctgatgca atagctgaaa taaaacaatt
     4261 taatcgattt gcccatgacc caatggctgg cggtcacaga atgtggcaaa tggccgggct
     4321 taaagcccag cgggcgcaga cggatgtaaa taataagcag gctgcatttg atgctgctgc
     4381 aaaagagaag tcagatgctg atgctgcatt gagttctgct atggaaagca ggaagaagaa
     4441 agaagataag aaaaggagtg ctgaaaataa tttaaacgat gaaaagaata agcccagaaa
     4501 aggttttaaa gattacgggc atgattatca tccagctccg aaaactgaga atattaaagg
     4561 gcttggtgat cttaagcctg ggataccaaa aacaccaaag cagaatggtg gtggaaaacg
     4621 caagcgctgg actggagata aagggcgtaa gatttatgag tgggattctc agcatggtga
     4681 gcttgagggg tatcgtgcca gtgatggtca gcatcttggc tcatttgacc ctaaaacagg
     4741 caatcagttg aaaggtccag atccgaaacg aaatatcaag aaatatcttt gagaattcga
     4801 gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttg cggccgcgtc
     4861 gtgactggga aaaccctggc gcaaagcccg ccgaaaggcg ggcttttctg ttataaacgc
     4921 agaaaggccc acccgaaggt gagccagtgt gactctagta gagagcgttc accgacaaaa
     4981 acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt gatgcctggc
     5041 tcgagctcga gaaaaaaagc ccggacgact gttcgggctt gtctttttat atgttggaaa
     5101 atcagtggca atgcaatggc ccaacagaaa atggtgaaac cagtaacgtt atactgaatc
     5161 taagtatcaa tgtgagcgct cacaagtcaa cactcttttt gataaaacta gtcttggact
     5221 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
     5281 ttgccgccgg gcgtttttta ttggtgagaa tccaggggtc cccaataatt acgatttacg
     5341 tatttaaatg aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg
     5401 cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc
     5461 gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg cagcagggca
     5521 gtcgccctaa aacaaagtta aacatcatga gggaagcggt gatcgccgaa gtatcgactc
     5581 aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg ctggccgtac
     5641 atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt gatttgctgg
     5701 ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg
     5761 aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc accattgttg
     5821 tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa tttggagaat
     5881 ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac attgatctgg
     5941 ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca gcggcggagg
     6001 aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa accttaacgc
     6061 tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc
     6121 gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg
     6181 caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga caggcttatc
     6241 ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa tttgtccact
     6301 acgtgaaagg cgagatcacc aaggtagtcg gcaaataaga caattgtcct tttccgctgc
     6361 ataaccctgc ttcggggtca ttatagcgat tttttcggta tatccatcct ttttcgcacg
     6421 atatacagga ttttgccaaa gggttcgtgt agactttcct tggtgtatcc aacggcgtca
     6481 gccgggcagg ataggtgaag taggcccacc cgcgagcggg tgttccttct tcactgtccc
     6541 ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg cgaggctggc cgtaggccgg
     6601 ccctaccggc gcggcagcgt tacccgtgtc ggcggctcca acggctcgcc atcgtccaga
     6661 aaacacggct catcgggcat cggcaggcgc tgctgcccgc gccgttccca ttcctccgtt
     6721 tcggtcaagg ctggcaggtc tggttccatg cccggaatgc cgggctggct gggcggctcc
     6781 tcgccggggc cggtcggtag ttgctgctcg cccggataca gggtcgggat gcggcgcagg
     6841 tcgccatgcc ccaacagcga ttcgtcctgg tcgtcgtgat caaccaccac ggcggcactg
     6901 aacaccgaca ggcgcaactg gtcgcggggc tggccccacg ccacgcggtc attgaccacg
     6961 taggccgaca cggtgccggg gccgttgagc ttcacgacgg agatccagcg ctcggccacc
     7021 aagtccttga ctgcgtattg gaccgtccgc aaagaacgtc cgatgagctt ggaaagtgtc
     7081 ttctggctga ccaccacggc gttctggtgg cccatctgcg ccacgaggtg atgcagcagc
     7141 attgccgccg tgggtttcct cgcaataagc ccggcccacg cctcatgcgc tttgcgttcc
     7201 gtttgcaccc agtgaccggg cttgttcttg gcttgaatgc cgatttctct ggactgcgtg
     7261 gccatgctta tctccatgcg gtaggggtgc cgcacggttg cggcaccatg cgcaatcagc
     7321 tgcaactttt cggcagcgcg acaacaatta tgcgttgcgt aaaagtggca gtcaattaca
     7381 gattttcttt aacctacgca atgagctatt gcggggggtg ccgcaatgag ctgttgcgta
     7441 cccccctttt ttaagttgtt gatttttaag tctttcgcat ttcgccctat atctagttct
     7501 ttggtgccca aagaagggca cccctgcggg gttcccccac gccttcggcg cggctccccc
     7561 tccggcaaaa agtggcccct ccggggcttg ttgatcgact gcgcggcctt cggccttgcc
     7621 caaggtggcg ctgccccctt ggaacccccg cactcgccgc cgtgaggctc ggggggcagg
     7681 cgggcgggct tcgcccttcg actgccccca ctcgcatagg cttgggtcgt tccaggcgcg
     7741 tcaaggccaa gccgctgcgc ggtcgctgcg cgagccttga cccgccttcc acttggtgtc
     7801 caaccggcaa gcgaagcgcg caggccgcag gccggaggct tttccccaga gaaaattaaa
     7861 aaaattgatg gggcaaggcc gcaggccgcg cagttggagc cggtgggtat gtggtcgaag
     7921 gctgggtagc cggtgggcaa tccctgtggt caagctcgtg ggcaggcgca gcctgtccat
     7981 cagcttgtcc agcagggttg tccacgggcc gagcgaagcg agccagccgg tggccgctcg
     8041 cggccatcgt ccacatatcc acgggctggc aagggagcgc agcgaccgcg cagggcgaag
     8101 cccggagagc aagcccgtag ggggggcgcg cccagctgtc tagggcggcg gatttgtcct
     8161 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg
     8221 agcctttcgt tttatttgat gcct
//
