LOCUS       V008435                10136 bp    DNA     circular SYN 17-DEC-2018
DEFINITION  Exported.
ACCESSION   V008435
VERSION     V008435
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10136)
  AUTHORS   Chou H-H., Berthet J, Marx CJ.
  TITLE     Growth rate modulates the selective advantage of recurrent
            transposition mutations arising from experimental populations grown
            in a metal-limited environment
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10136)
  AUTHORS   Marx CJ, Chou H-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-OCT-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138,
            USA
REFERENCE   3  (bases 1 to 10136)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10136)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (15-OCT-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138,
            USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10136
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..712
                     /label="oriV"
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    1981..2002
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2017..2047
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2055..2071
                     /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)."
     primer_bind     2079..2095
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     regulatory      2206..2306
                     /label="PmxaF"
                     /note="PmxaF"
                     /regulatory_class="promoter"
     primer_bind     2446..2462
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    complement(2475..2531)
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     CDS             2622..3746
                     /gene="flhA"
                     /label="S-(hydroxymethyl)glutathione dehydrogenase"
                     /note="S-(hydroxymethyl)glutathione dehydrogenase from
                     Paracoccus denitrificans. Accession#: P45382"
     CDS             3803..4642
                     /codon_start=1
                     /gene="fghA"
                     /product="S-formylglutathione hydrolase"
                     /label="fghA"
                     /note="S"
                     /protein_id="ACR45017.1"
                     /translation="MTLAYETVSENRSFGGIQGVYRHQSQATGTPMTFAIYLPPDARHG
                     KVPVLWYLSGLTCTHENAMTKAGAQEWAAEYGIAVIFPDTSPRGEGVANDETYDLGQGA
                     GFYVDATEAPWAPHFRMWHYVTHELPELVFNNFPLDREAQGITGHSMGGHGALTIAMTF
                     PERYRSVSAFAPIAHPSESDWGRKQFAAYLGDDKAAWKRHDSTILMREKGYPGEVLIDQ
                     GASDQFLDLLKPEALAHAMAERRQPGTFRMQQGYDHSYFFVQSFMADHIRWHAERLG"
     gene            3803..4642
                     /gene="fghA"
                     /label="fghA"
     primer_bind     complement(4694..4710)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     CDS             complement(5155..5796)
                     /label="TetR"
                     /note="tetracycline resistance regulatory protein"
     CDS             complement(6577..7389)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(8064..9209)
                     /label="trfA"
                     /note="trans-acting replication protein that binds to and
                     activates oriV"
     CDS             complement(9481..9852)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label="traJ"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(9885..9994)
                     /direction=LEFT
                     /label="oriT"
                     /note="incP origin of transfer"
ORIGIN
        1 gaccctttcc gacgctcacc gggctggttg ccctcgccgc tgggctggcg gccgtctatg
       61 gccctgcaaa cgcgccagaa acgccgtcga agccgtgtgc gagacaccgc ggccgccggc
      121 gttgtggata cctcgcggaa aacttggccc tcactgacag atgaggggcg gacgttgaca
      181 cttgaggggc cgactcaccc ggcgcggcgt tgacagatga ggggcaggct cgatttcggc
      241 cggcgacgtg gagctggcca gcctcgcaaa tcggcgaaaa cgcctgattt tacgcgagtt
      301 tcccacagat gatgtggaca agcctgggga taagtgccct gcggtattga cacttgaggg
      361 gcgcgactac tgacagatga ggggcgcgat ccttgacact tgaggggcag agtgctgaca
      421 gatgaggggc gcacctattg acatttgagg ggctgtccac aggcagaaaa tccagcattt
      481 gcaagggttt ccgcccgttt ttcggccacc gctaacctgt cttttaacct gcttttaaac
      541 caatatttat aaaccttgtt tttaaccagg gctgcgccct gtgcgcgtga ccgcgcacgc
      601 cgaagggggg tgccccccct tctcgaaccc tcccggcccg ctaacgcggg cctcccatcc
      661 ccccaggggc tgcgcccctc ggccgcgaac ggcctcaccc caaaaatggc agccaagctg
      721 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
      781 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
      841 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
      901 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
      961 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     1021 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     1081 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     1141 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     1201 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     1261 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     1321 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     1381 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     1441 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     1501 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     1561 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     1621 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     1681 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     1741 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     1801 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     1861 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     1921 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     1981 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     2041 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     2101 ttacgccaag ctagcttccc gcttggtcgg gccgcttcgc gagggcccgt tgacgacaac
     2161 ggtgcgatgg gtcccggccc cggtcaagac gatgccaata cgttgcgaca ctacgccttg
     2221 gcacttttag aattgcctta tcgtcctgat aagaaatgtc cgaccagcta aagacatcgc
     2281 gtccaatcaa agcctagaaa atataggcga agggacgcta ataagtcttt cataagaccg
     2341 cgcaaatcta aaaatatcct tagattcacg atgcggcact tcggatgact tccgagcgag
     2401 cctggaacct cagaaaaacg tctgagagat accgcggatc tcacacagga aacagctatg
     2461 accatgatta cgccaagctt gcatgcctgc aggtcgactc tagaggatcc ccgggtaccg
     2521 agctcgaatt cgcccttgtc tagatgcatg ctcgagcggc cgccagtgtg atggatatct
     2581 gcagaattcg cccttgaggc ctacatgaag gagaagacaa gatgagaacc cgtgccgccg
     2641 tggcccttga ggctggcaaa ccgctggaag tgatggaggt caacctggaa ggcccgaagg
     2701 ccggcgaggt gatggtcgag atcaaggcca ccggcatctg ccacaccgac gaattcaccc
     2761 tgtcgggcgc cgatcccgag gggctcttcc cctcgatcct gggccatgag ggcgcgggcg
     2821 tcgtggtcga ggtcggcccg ggcgttacct cggtcaagcc gggcgatcat gtcatcccgc
     2881 tctacacgcc cgagtgccgg caatgcgcgt cctgcctgtc gggcaagacc aacctttgca
     2941 ccgccattcg cgccacccag ggtcagggcc tgatgccgga cggaaccagc cggttcagca
     3001 tgctcgacgg cacgccgatc tttcattaca tgggctgctc gaccttctcg aactacaccg
     3061 tgctgccgga aatcgcggtg gccaaggtgc gcgaggacgc gcccttcgac aagatctgct
     3121 atatcggctg cggcgtcact accggcatcg gcgcggtgat caacaccgcc aaggtggaaa
     3181 tcggcgccaa ggcggtggtc ttcggcctgg gcggcattgg gctgaacgtg ctgcaggggc
     3241 tgcgcctggc cggcgcggac atgatcatcg gcgtcgatct gaacgacgac aagaagccga
     3301 tggccgagca tttcggcatg acgcatttca tcaacccgaa gaattgcgag aacgtggtgc
     3361 aggagatcgt gaacctgacc aagacgccct tcgaccagat cggcggcgcg gattacagct
     3421 tcgactgcac cggcaacgtc aaggtgatgc gcgacgcgct ggaatgcacc catcgcggct
     3481 ggggccagtc gatcatcatc ggcgtggcgc cggcgggggc cgagatcagc acccgcccgt
     3541 tccagctggt caccggccgg gtctggaagg gcacggcctt cggcggtgcc cgcggccgca
     3601 ccgacgtgcc gcagatcgtc gactggtata tggacggcaa gatcgagatc gacccgatga
     3661 tcacccacac cctgtcgctg gacgacatca acaagggctt cgacctgatg catgcgggcg
     3721 aatccatccg ctcggtcgtc ctttactgat cgtcaagggc tgcttatcta gagttccacg
     3781 acttgacaga agggaagaac ccatgacgct ggcctatgaa accgtatcgg aaaaccgcag
     3841 cttcggcggt atccagggcg tctatcgcca ccagtcccag gcgaccggca cgccgatgac
     3901 ctttgccatc tacctgccgc ccgatgcccg gcacggcaag gtgccggtgc tgtggtatct
     3961 ttcgggcctg acctgcacgc atgaaaatgc catgaccaag gccggcgcac aggaatgggc
     4021 ggcggaatac ggcatcgcgg tgatctttcc cgacacctcg ccgcgcggcg aaggcgtggc
     4081 caacgacgag acctacgacc tgggccaggg cgccggcttc tatgtcgatg cgaccgaggc
     4141 gccctgggcg ccgcatttca ggatgtggca ctatgtcacc catgaactgc ccgagctggt
     4201 cttcaacaac ttcccgctgg accgcgaggc gcagggcatc accggccatt ccatgggcgg
     4261 gcacggtgcg ctgaccatcg ccatgacctt cccggagcgc tatcgctcgg tctcggcctt
     4321 tgcgccgatc gcgcatccca gcgaatccga ctgggggcgc aaacagttcg ccgcctatct
     4381 gggcgacgac aaggcggcat ggaaaaggca tgattcgacc atcctgatgc gcgaaaaggg
     4441 ctatcccggc gaggtgctga tcgaccaggg cgcgagcgac cagttcctcg atcttctgaa
     4501 gcccgaggcg ctggcccatg ccatggccga acgccgccag cccggcacct tccgcatgca
     4561 gcagggctac gatcacagct atttcttcgt gcaaagcttc atggccgacc atatccgctg
     4621 gcacgcggag cggctgggct aggcggccgg gaccggccaa agggcaacgg ggcagagctc
     4681 aaagggcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     4741 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     4801 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     4861 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     4921 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4981 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     5041 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     5101 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagcaccctt
     5161 tctcggtcct tcaacgttcc tgacaacgag cctccttttc gccaatccat cgacaatcac
     5221 cgcgagtccc tgctcgaacg ctgcgtccgg accggcttcg tcgaaggcgt ctatcgcggc
     5281 ccgcaacagc ggcgagagcg gagcctgttc aacggtgccg ccgcgctcgc cggcatcgct
     5341 gtcgccggcc tgctcctcaa gcacggcccc aacagtgaag tagctgattg tcatcagcgc
     5401 attgacggcg tccccggccg aaaaacccgc ctcgcagagg aagcgaagct gcgcgtcggc
     5461 cgtttccatc tgcggtgcgc ccggtcgcgt gccggcatgg atgcgcgcgc catcgcggta
     5521 ggcgagcagc gcctgcctga agctgcgggc attcccgatc agaaatgagc gccagtcgtc
     5581 gtcggctctc ggcaccgaat gcgtatgatt ctccgccagc atggcttcgg ccagtgcgtc
     5641 gagcagcgcc cgcttgttcc tgaagtgcca gtaaagcgcc ggctgctgaa cccccaaccg
     5701 ttccgccagt ttgcgtgtcg tcagaccgtc tacgccgacc tcgttcaaca ggtccagggc
     5761 ggcacggatc actgtattcg gctgcaactt tgtcatgctt gacactttat cactgataaa
     5821 cataatatgt ccaccaactt atcagtgata aagaatccgc gcgttcaatc ggaccagcgg
     5881 aggctggtcc ggaggccaga cgtgaaaccc aacatacccc tgatcgtaat tctgagcact
     5941 gtcgcgctcg acgctgtcgg catcggcctg attatgccgg tgctgccggg cctcctgcgc
     6001 gatctggttc actcgaacga cgtcaccgcc cactatggca ttctgctggc gctgtatgcg
     6061 ttggtgcaat ttgcctgcgc acctgtgctg ggcgcgctgt cggatcgttt cgggcggcgg
     6121 ccaatcttgc tcgtctcgct ggccggcgcc actgtcgact acgccatcat ggcgacagcg
     6181 cctttccttt gggttctcta tatcgggcgg atcgtggccg gcatcaccgg ggcgactggg
     6241 gcggtagccg gcgcttatat tgccgatgac ctgcaggggg gggggggcgc tgaggtctgc
     6301 ctcgtgaaga aggtgttgct gactcatacc aggcctgaat cgccccatca tccagccaga
     6361 aagtgaggga gccacggttg atgagagctt tgttgtaggt ggaccagttg gtgattttga
     6421 acttttgctt tgccacggaa cggtctgcgt tgtcgggaag atgcgtgatc tgatccttca
     6481 actcagcaaa agttcgattt attcaacaaa gccgccgtcc cgtcaagtca gcgtaatgct
     6541 ctgccagtgt tacaaccaat taaccaattc tgattagaaa aactcatcga gcatcaaatg
     6601 aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa gccgtttctg
     6661 taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct ggtatcggtc
     6721 tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt caaaaataag
     6781 gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg gcaaaagctt
     6841 atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat caaaatcact
     6901 cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc
     6961 gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga acactgccag
     7021 cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga atgctgtttt
     7081 cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa aatgcttgat
     7141 ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat ctgtaacatc
     7201 attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg gcttcccata
     7261 caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt tatacccata
     7321 taaatcagca tccatgttgg aatttaatcg cggcctcgag caagacgttt cccgttgaat
     7381 atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta ttgttcatga
     7441 tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa cgtggctttc
     7501 cccccccccc ctgcaggtcc gacacgggga tggatggcgt tcccgatcat ggtcctgctt
     7561 gcttcgggtg gcatcggaat gccggcgctg caagcaatgt tgtccaggca ggtggatgag
     7621 gaacgtcagg ggcagctgca aggctcactg gcggcgctca ccagcctgac ctcgatcgtc
     7681 ggacccctcc tcttcacggc gatctatgcg gcttctataa caacgtggaa cgggtgggca
     7741 tggattgcag gcgctgccct ctacttgctc tgcctgccgg cgctgcgtcg cgggctttgg
     7801 agcggcgcag ggcaacgagc cgatcgctga tcgtggaaac gataggccta tgccatgcgg
     7861 gtcaaggcga cttccggcaa gctatacgcg ccctagaatt gtcaatttta atcctctgtt
     7921 tatcggcagt tcgtagagcg cgccgtgcgt cccgagcgat actgagcgaa gcaagtgcgt
     7981 cgagcagtgc ccgcttgttc ctgaaatgcc agtaaagcgc tggctgctga acccccagcc
     8041 ggaactgacc ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg
     8101 tgttccacca ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac
     8161 ttcttcacgc gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg
     8221 tacggctccc ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg
     8281 cggtacttct cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc
     8341 tcgtcgatca ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg
     8401 tgcagcagcg acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc
     8461 gcctgtaggc gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag
     8521 cccaggtcct ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc
     8581 ttcgcgtact ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg
     8641 ccggtgtagg tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc
     8701 tcgcgcggga ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc
     8761 atcgctcgca tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg
     8821 atctgctgct tcgtgtgttt cagcaacgcg gcctgcttgg cctcgctgac ctgttttgcc
     8881 aggtcctcgc cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc
     8941 atcgacttcg ccaaacctgc cgcctcctgt tcgagacgac gcgaacgctc cacggcggcc
     9001 gatggcgcgg gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta
     9061 gcttgctgga ccatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg
     9121 cggcttgcga tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg
     9181 tatgccttcc ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccgactcac
     9241 gccggggcaa tgtgccctta ttcctgattt gacccgcctg gtgccttggt gtccagataa
     9301 tccaccttat cggcaatgaa gtcggtcccg tagaccgtct ggccgtcctt ctcgtacttg
     9361 gtattccgaa tcttgccctg cacgaatacc agctccgcga agtcgctctt cttgatggag
     9421 cgcatgggga cgtgcttggc aatcacgcgc accccccggc cgttttagcg gctaaaaaag
     9481 tcatggctct gccctcgggc ggaccacgcc catcatgacc ttgccaagct cgtcctgctt
     9541 ctcttcgatc ttcgccagca gggcgaggat cgtggcatca ccgaaccgcg ccgtgcgcgg
     9601 gtcgtcggtg agccagagtt tcagcaggcc gcccaggcgg cccaggtcgc cattgatgcg
     9661 ggccagctcg cggacgtgct catagtccac gacgcccgtg attttgtagc cctggccgac
     9721 ggccagcagg taggcctaca ggctcatgcc ggccgccgcc gccttttcct caatcgctct
     9781 tcgttcgtct ggaaggcagt acaccttgat aggtgggctg cccttcctgg ttggcttggt
     9841 ttcatcagcc atccgcttgc cctcatctgt tacgccggcg gtagccggcc agcctcgcag
     9901 agcaggattc ccgttgagca ccgccaggtg cgaataaggg acagtgaaga aggaacaccc
     9961 gctcgcgggt gggcctactt cacctatcct gcccggctga cgccgttgga tacaccaagg
    10021 aaagtctaca cgaacccttt ggcaaaatcc tgtatatcgt gcgaaaaagg atggatatac
    10081 cgaaaaaatc gctataatga ccccgaagca gggttatgca gcggaaaaga tccgtc
//
