LOCUS       40924_24218        9129 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC46, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9129)
  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  JOURNAL   Unpublished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 9129)
  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 9129)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9129)
  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            "Unpublished"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..9129
                     /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"
     misc_feature    2920..3201
                     /label=5' upstream region of icuAB.1145
                     /note="5' upstream region of icuAB.1145"
     terminator      complement(3300..3327)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3419..3505)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(3687..3703)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(4148..4789)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKG"
     CDS             complement(5570..6382)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(7057..8202)
                     /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(8474..8845)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(8878..8987)
                     /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 gtgcaaccta ggcactcagt
     2941 gatccgggcg ctactaaact gcaaattatg ggcaggcaca agataggaat ttttcgtatt
     3001 cgtacgtctc atcgagatgt tgcagccttt agcccgtctt attcacgagg gcagtggatc
     3061 tgtggtggcg agcgtagcgt aaagcattga tgtggcttag ggattgggtg ctgacgccaa
     3121 gtcctcagac cagatcgagc ccgccacgcc cgccatgtcc gatcctacgt gtctgccgtt
     3181 cgcgtttcca tcggttcgag gaattccctg ctttcctgat gcaaaaacga ggctagttta
     3241 ccgtatctgt ggggggatgg cttgtagata tgacgacagg aagagtttgt agaaacgcaa
     3301 aaaggccatc cgtcaggatg gccttctgct taatttgatg cctggcagtt tatggcgggc
     3361 gtcctgcccg ccaccctccg ggccgttgct tcgcaacgtt caaatccgct cccggcggat
     3421 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
     3481 tcgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc atggggagac
     3541 cccacactac catcggcgct acggcgtttc acttctgagt tcggcatggg gtcaggtggg
     3601 accaccgcgc tactgccgcc aggcaaattc tgttttatca gaccgcttct gcgttctgat
     3661 ttaatctgta tcaggctgaa aaattcactg gccgtcgttt tacaacgtcg tgactgggaa
     3721 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
     3781 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     3841 tggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     3901 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     3961 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4021 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4081 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     4141 tcttagcacc ctttctcggt ccttcaacgt tcctgacaac gagcctcctt ttcgccaatc
     4201 catcgacaat caccgcgagt ccctgctcga acgctgcgtc cggaccggct tcgtcgaagg
     4261 cgtctatcgc ggcccgcaac agcggcgaga gcggagcctg ttcaacggtg ccgccgcgct
     4321 cgccggcatc gctgtcgccg gcctgctcct caagcacggc cccaacagtg aagtagctga
     4381 ttgtcatcag cgcattgacg gcgtccccgg ccgaaaaacc cgcctcgcag aggaagcgaa
     4441 gctgcgcgtc ggccgtttcc atctgcggtg cgcccggtcg cgtgccggca tggatgcgcg
     4501 cgccatcgcg gtaggcgagc agcgcctgcc tgaagctgcg ggcattcccg atcagaaatg
     4561 agcgccagtc gtcgtcggct ctcggcaccg aatgcgtatg attctccgcc agcatggctt
     4621 cggccagtgc gtcgagcagc gcccgcttgt tcctgaagtg ccagtaaagc gccggctgct
     4681 gaacccccaa ccgttccgcc agtttgcgtg tcgtcagacc gtctacgccg acctcgttca
     4741 acaggtccag ggcggcacgg atcactgtat tcggctgcaa ctttgtcatg attgacactt
     4801 tatcactgat aaacataata tgtccaccaa cttatcagtg ataaagaatc cgcgcgttca
     4861 atcggaccag cggaggctgg tccggaggcc agacgtgaaa cccaacatac ccctgatcgt
     4921 aattctgagc actgtcgcgc tcgacgctgt cggcatcggc ctgattatgc cggtgctgcc
     4981 gggcctcctg cgcgatctgg ttcactcgaa cgacgtcacc gcccactatg gcattctgct
     5041 ggcgctgtat gcgttggtgc aatttgcctg cgcacctgtg ctgggcgcgc tgtcggatcg
     5101 tttcgggcgg cggccaatct tgctcgtctc gctggccggc gccactgtcg actacgccat
     5161 catggcgaca gcgcctttcc tttgggttct ctatatcggg cggatcgtgg ccggcatcac
     5221 cggggcgact ggggcggtag ccggcgctta tattgccgat gacctgcagg gggggggggg
     5281 cgctgaggtc tgcctcgtga agaaggtgtt gctgactcat accaggcctg aatcgcccca
     5341 tcatccagcc agaaagtgag ggagccacgg ttgatgagag ctttgttgta ggtggaccag
     5401 ttggtgattt tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg
     5461 atctgatcct tcaactcagc aaaagttcga tttattcaac aaagccgccg tcccgtcaag
     5521 tcagcgtaat gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat
     5581 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     5641 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     5701 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     5761 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     5821 atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     5881 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     5941 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     6001 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     6061 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     6121 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     6181 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     6241 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     6301 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctc gagcaagacg
     6361 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     6421 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     6481 caacgtggct ttcccccccc cccctgcagg tccgacacgg ggatggatgg cgttcccgat
     6541 catggtcctg cttgcttcgg gtggcatcgg aatgccggcg ctgcaagcaa tgttgtccag
     6601 gcaggtggat gaggaacgtc aggggcagct gcaaggctca ctggcggcgc tcaccagcct
     6661 gacctcgatc gtcggacccc tcctcttcac ggcgatctat gcggcttcta taacaacgtg
     6721 gaacgggtgg gcatggattg caggcgctgc cctctacttg ctctgcctgc cggcgctgcg
     6781 tcgcgggctt tggagcggcg cagggcaacg agccgatcgc tgatcgtgga aacgataggc
     6841 ctatgccatg cgggtcaagg cgacttccgg caagctatac gcgccctaga attgtcaatt
     6901 ttaatcctct gtttatcggc agttcgtaga gcgcgccgtg cgtcccgagc gatactgagc
     6961 gaagcaagtg cgtcgagcag tgcccgcttg ttcctgaaat gccagtaaag cgctggctgc
     7021 tgaaccccca gccggaactg accccacaag gccctagcgt ttgcaatgca ccaggtcatc
     7081 attgacccag gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac
     7141 ctgctcgcgc cacttcttca cgcgggtgga atccgatccg cacatgaggc ggaaggtttc
     7201 cagcttgagc gggtacggct cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt
     7261 cggcgacagc ttgcggtact tctcccatat gaatttcgtg tagtggtcgc cagcaaacag
     7321 cacgacgatt tcctcgtcga tcaggacctg gcaacgggac gttttcttgc cacggtccag
     7381 gacgcggaag cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa
     7441 gcccatcgcc gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt aataccggcc
     7501 attgatcgac cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc
     7561 gataggggtg cgcttcgcgt actccaacac ctgctgccac accagttcgt catcgtcggc
     7621 ccgcagctcg acgccggtgt aggtgatctt cacgtccttg ttgacgtgga aaatgacctt
     7681 gttttgcagc gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc
     7741 cgtgtcgttt ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga acaaggaaag
     7801 ctgcatttcc ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct
     7861 gacctgtttt gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg
     7921 cgtgtcgatg gtcatcgact tcgccaaacc tgccgcctcc tgttcgagac gacgcgaacg
     7981 ctccacggcg gccgatggcg cgggcagggc agggggagcc agttgcacgc tgtcgcgctc
     8041 gatcttggcc gtagcttgct ggaccatcga gccgacggac tggaaggttt cgcggggcgc
     8101 acgcatgacg gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat
     8161 cagttcttgc ctgtatgcct tccggtcaaa cgtccgattc attcaccctc cttgcgggat
     8221 tgccccgact cacgccgggg caatgtgccc ttattcctga tttgacccgc ctggtgcctt
     8281 ggtgtccaga taatccacct tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc
     8341 cttctcgtac ttggtattcc gaatcttgcc ctgcacgaat accagctccg cgaagtcgct
     8401 cttcttgatg gagcgcatgg ggacgtgctt ggcaatcacg cgcacccccc ggccgtttta
     8461 gcggctaaaa aagtcatggc tctgccctcg ggcggaccac gcccatcatg accttgccaa
     8521 gctcgtcctg cttctcttcg atcttcgcca gcagggcgag gatcgtggca tcaccgaacc
     8581 gcgccgtgcg cgggtcgtcg gtgagccaga gtttcagcag gccgcccagg cggcccaggt
     8641 cgccattgat gcgggccagc tcgcggacgt gctcatagtc cacgacgccc gtgattttgt
     8701 agccctggcc gacggccagc aggtaggcct acaggctcat gccggccgcc gccgcctttt
     8761 cctcaatcgc tcttcgttcg tctggaaggc agtacacctt gataggtggg ctgcccttcc
     8821 tggttggctt ggtttcatca gccatccgct tgccctcatc tgttacgccg gcggtagccg
     8881 gccagcctcg cagagcagga ttcccgttga gcaccgccag gtgcgaataa gggacagtga
     8941 agaaggaaca cccgctcgcg ggtgggccta cttcacctat cctgcccggc tgacgccgtt
     9001 ggatacacca aggaaagtct acacgaaccc tttggcaaaa tcctgtatat cgtgcgaaaa
     9061 aggatggata taccgaaaaa atcgctataa tgaccccgaa gcagggttat gcagcggaaa
     9121 agatccgtc
//
