LOCUS       40924_24243        8898 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC62, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8898)
  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  JOURNAL   Unpublished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 8898)
  AUTHORS   Chou H-H.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-OCT-2008) Organismic and Evolutionary Biology, Harvard
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA
REFERENCE   3  (bases 1 to 8898)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8898)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (14-OCT-2008) Organismic and Evolutionary Biology, Harvard 
            University, 16 Divinity Avenue, Biolab 3079, Cambridge, MA 02138, 
            USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8898
                     /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
                     /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"
     CDS             complement(2123..2836)
                     /codon_start=1
                     /label=GFPuv
                     /note="GFP variant optimized for excitation by UV light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     promoter        complement(2927..2955)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     terminator      complement(3069..3096)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3188..3274)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(3456..3472)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3917..4558)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKG"
     CDS             complement(5339..6151)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6826..7971)
                     /codon_start=1
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(8243..8614)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(8647..8756)
                     /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 tccttctagt
     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 cttgcatgct tatttgtaga gctcatccat gccatgtgta atcccagcag
     2161 cagttacaaa ctcaagaagg accatgtggt cacgcttttc gttgggatct ttcgaaaggg
     2221 cagattgtgt cgacaggtaa tggttgtctg gtaaaaggac agggccatcg ccaattggag
     2281 tattttgttg ataatggtct gctagttgaa cggatccatc ttcaatgttg tggcgaattt
     2341 tgaagttagc tttgattcca ttcttttgtt tgtctgccgt gatgtataca ttgtgtgagt
     2401 tatagttgta ctcgagtttg tgtccgagaa tgtttccatc ttctttaaaa tcaatacctt
     2461 ttaactcgat acgattaaca agggtatcac cttcaaactt gacttcagca cgcgtcttgt
     2521 agttcccgtc atctttgaaa gatatagtgc gttcctgtac ataaccttcg ggcatggcac
     2581 tcttgaaaaa gtcatgccgt ttcatatgat ccggataacg ggaaaagcat tgaacaccat
     2641 aagagaaagt agtgacaagt gttggccatg gaacaggtag ttttccagta gtgcaaataa
     2701 atttaagggt aagttttccg tatgttgcat caccttcacc ctctccactg acagaaaatt
     2761 tgtgcccatt aacatcacca tctaattcaa caagaattgg gacaactcca gtgaaaagtt
     2821 cttctccttt actcattcgg ggtctctccc tgctagcggg cccggatccg atatcttaag
     2881 atcttccgtg cagtttaagc cgaattgatc cccgggtacc tccacacatt atacgagccg
     2941 atgattaatt gtcaacagct catttcagag aattccctgc tttcctgatg caaaaacgag
     3001 gctagtttac cgtatctgtg gggggatggc ttgtagatat gacgacagga agagtttgta
     3061 gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc ctggcagttt
     3121 atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc aaatccgctc
     3181 ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata aaacgaaagg
     3241 cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca
     3301 tggggagacc ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg
     3361 tcaggtggga ccaccgcgct actgccgcca ggcaaattct gttttatcag accgcttctg
     3421 cgttctgatt taatctgtat caggctgaaa aattcactgg ccgtcgtttt acaacgtcgt
     3481 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
     3541 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
     3601 aatggcgaat ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     3661 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga
     3721 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     3781 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     3841 aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3901 ataatggttt cttagcaccc tttctcggtc cttcaacgtt cctgacaacg agcctccttt
     3961 tcgccaatcc atcgacaatc accgcgagtc cctgctcgaa cgctgcgtcc ggaccggctt
     4021 cgtcgaaggc gtctatcgcg gcccgcaaca gcggcgagag cggagcctgt tcaacggtgc
     4081 cgccgcgctc gccggcatcg ctgtcgccgg cctgctcctc aagcacggcc ccaacagtga
     4141 agtagctgat tgtcatcagc gcattgacgg cgtccccggc cgaaaaaccc gcctcgcaga
     4201 ggaagcgaag ctgcgcgtcg gccgtttcca tctgcggtgc gcccggtcgc gtgccggcat
     4261 ggatgcgcgc gccatcgcgg taggcgagca gcgcctgcct gaagctgcgg gcattcccga
     4321 tcagaaatga gcgccagtcg tcgtcggctc tcggcaccga atgcgtatga ttctccgcca
     4381 gcatggcttc ggccagtgcg tcgagcagcg cccgcttgtt cctgaagtgc cagtaaagcg
     4441 ccggctgctg aacccccaac cgttccgcca gtttgcgtgt cgtcagaccg tctacgccga
     4501 cctcgttcaa caggtccagg gcggcacgga tcactgtatt cggctgcaac tttgtcatga
     4561 ttgacacttt atcactgata aacataatat gtccaccaac ttatcagtga taaagaatcc
     4621 gcgcgttcaa tcggaccagc ggaggctggt ccggaggcca gacgtgaaac ccaacatacc
     4681 cctgatcgta attctgagca ctgtcgcgct cgacgctgtc ggcatcggcc tgattatgcc
     4741 ggtgctgccg ggcctcctgc gcgatctggt tcactcgaac gacgtcaccg cccactatgg
     4801 cattctgctg gcgctgtatg cgttggtgca atttgcctgc gcacctgtgc tgggcgcgct
     4861 gtcggatcgt ttcgggcggc ggccaatctt gctcgtctcg ctggccggcg ccactgtcga
     4921 ctacgccatc atggcgacag cgcctttcct ttgggttctc tatatcgggc ggatcgtggc
     4981 cggcatcacc ggggcgactg gggcggtagc cggcgcttat attgccgatg acctgcaggg
     5041 gggggggggc gctgaggtct gcctcgtgaa gaaggtgttg ctgactcata ccaggcctga
     5101 atcgccccat catccagcca gaaagtgagg gagccacggt tgatgagagc tttgttgtag
     5161 gtggaccagt tggtgatttt gaacttttgc tttgccacgg aacggtctgc gttgtcggga
     5221 agatgcgtga tctgatcctt caactcagca aaagttcgat ttattcaaca aagccgccgt
     5281 cccgtcaagt cagcgtaatg ctctgccagt gttacaacca attaaccaat tctgattaga
     5341 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     5401 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     5461 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     5521 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     5581 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     5641 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     5701 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     5761 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     5821 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     5881 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     5941 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     6001 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     6061 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctcg
     6121 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     6181 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     6241 tttgagacac aacgtggctt tccccccccc ccctgcaggt ccgacacggg gatggatggc
     6301 gttcccgatc atggtcctgc ttgcttcggg tggcatcgga atgccggcgc tgcaagcaat
     6361 gttgtccagg caggtggatg aggaacgtca ggggcagctg caaggctcac tggcggcgct
     6421 caccagcctg acctcgatcg tcggacccct cctcttcacg gcgatctatg cggcttctat
     6481 aacaacgtgg aacgggtggg catggattgc aggcgctgcc ctctacttgc tctgcctgcc
     6541 ggcgctgcgt cgcgggcttt ggagcggcgc agggcaacga gccgatcgct gatcgtggaa
     6601 acgataggcc tatgccatgc gggtcaaggc gacttccggc aagctatacg cgccctagaa
     6661 ttgtcaattt taatcctctg tttatcggca gttcgtagag cgcgccgtgc gtcccgagcg
     6721 atactgagcg aagcaagtgc gtcgagcagt gcccgcttgt tcctgaaatg ccagtaaagc
     6781 gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac
     6841 caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc
     6901 ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg
     6961 gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg
     7021 tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc
     7081 agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc
     7141 acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc
     7201 ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta
     7261 ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat
     7321 cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc
     7381 atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa
     7441 aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc
     7501 agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa
     7561 caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt
     7621 ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat
     7681 agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcgagacg
     7741 acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct
     7801 gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact ggaaggtttc
     7861 gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc
     7921 cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc
     7981 ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat ttgacccgcc
     8041 tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc cgtagaccgt
     8101 ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata ccagctccgc
     8161 gaagtcgctc ttcttgatgg agcgcatggg gacgtgcttg gcaatcacgc gcaccccccg
     8221 gccgttttag cggctaaaaa agtcatggct ctgccctcgg gcggaccacg cccatcatga
     8281 ccttgccaag ctcgtcctgc ttctcttcga tcttcgccag cagggcgagg atcgtggcat
     8341 caccgaaccg cgccgtgcgc gggtcgtcgg tgagccagag tttcagcagg ccgcccaggc
     8401 ggcccaggtc gccattgatg cgggccagct cgcggacgtg ctcatagtcc acgacgcccg
     8461 tgattttgta gccctggccg acggccagca ggtaggccta caggctcatg ccggccgccg
     8521 ccgccttttc ctcaatcgct cttcgttcgt ctggaaggca gtacaccttg ataggtgggc
     8581 tgcccttcct ggttggcttg gtttcatcag ccatccgctt gccctcatct gttacgccgg
     8641 cggtagccgg ccagcctcgc agagcaggat tcccgttgag caccgccagg tgcgaataag
     8701 ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac ttcacctatc ctgcccggct
     8761 gacgccgttg gatacaccaa ggaaagtcta cacgaaccct ttggcaaaat cctgtatatc
     8821 gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat gaccccgaag cagggttatg
     8881 cagcggaaaa gatccgtc
//
