LOCUS       40924_24233        8967 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC55, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8967)
  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  JOURNAL   Unpublished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 8967)
  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 8967)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8967)
  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            "Unpublished"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..8967
                     /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    complement(2921..3038)
                     /label=5' upstream region of icuAB.1145
                     /note="5' upstream region of icuAB.1145"
     terminator      complement(3138..3165)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3257..3343)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(3525..3541)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3986..4627)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKG"
     CDS             complement(5408..6220)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6895..8040)
                     /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(8312..8683)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(8716..8825)
                     /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 cgtgcaacct aggcactcag
     2941 tgatccgggc gctactaaac tgcaaattat gggcaggcac aagataggaa tttttcgtat
     3001 tcgtacgtct catcgagatg ttgcagcctt tagcccgtga attccctgct ttcctgatgc
     3061 aaaaacgagg ctagtttacc gtatctgtgg ggggatggct tgtagatatg acgacaggaa
     3121 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc
     3181 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca
     3241 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     3301 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     3361 actctcgcat ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc
     3421 ggcatggggt caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcaga
     3481 ccgcttctgc gttctgattt aatctgtatc aggctgaaaa attcactggc cgtcgtttta
     3541 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     3601 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     3661 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     3721 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     3781 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3841 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3901 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3961 gtcatgataa taatggtttc ttagcaccct ttctcggtcc ttcaacgttc ctgacaacga
     4021 gcctcctttt cgccaatcca tcgacaatca ccgcgagtcc ctgctcgaac gctgcgtccg
     4081 gaccggcttc gtcgaaggcg tctatcgcgg cccgcaacag cggcgagagc ggagcctgtt
     4141 caacggtgcc gccgcgctcg ccggcatcgc tgtcgccggc ctgctcctca agcacggccc
     4201 caacagtgaa gtagctgatt gtcatcagcg cattgacggc gtccccggcc gaaaaacccg
     4261 cctcgcagag gaagcgaagc tgcgcgtcgg ccgtttccat ctgcggtgcg cccggtcgcg
     4321 tgccggcatg gatgcgcgcg ccatcgcggt aggcgagcag cgcctgcctg aagctgcggg
     4381 cattcccgat cagaaatgag cgccagtcgt cgtcggctct cggcaccgaa tgcgtatgat
     4441 tctccgccag catggcttcg gccagtgcgt cgagcagcgc ccgcttgttc ctgaagtgcc
     4501 agtaaagcgc cggctgctga acccccaacc gttccgccag tttgcgtgtc gtcagaccgt
     4561 ctacgccgac ctcgttcaac aggtccaggg cggcacggat cactgtattc ggctgcaact
     4621 ttgtcatgat tgacacttta tcactgataa acataatatg tccaccaact tatcagtgat
     4681 aaagaatccg cgcgttcaat cggaccagcg gaggctggtc cggaggccag acgtgaaacc
     4741 caacataccc ctgatcgtaa ttctgagcac tgtcgcgctc gacgctgtcg gcatcggcct
     4801 gattatgccg gtgctgccgg gcctcctgcg cgatctggtt cactcgaacg acgtcaccgc
     4861 ccactatggc attctgctgg cgctgtatgc gttggtgcaa tttgcctgcg cacctgtgct
     4921 gggcgcgctg tcggatcgtt tcgggcggcg gccaatcttg ctcgtctcgc tggccggcgc
     4981 cactgtcgac tacgccatca tggcgacagc gcctttcctt tgggttctct atatcgggcg
     5041 gatcgtggcc ggcatcaccg gggcgactgg ggcggtagcc ggcgcttata ttgccgatga
     5101 cctgcagggg ggggggggcg ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac
     5161 caggcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct
     5221 ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg
     5281 ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa
     5341 agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     5401 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     5461 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     5521 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     5581 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     5641 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     5701 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     5761 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     5821 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     5881 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     5941 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     6001 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     6061 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     6121 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     6181 gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     6241 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     6301 atcagagatt ttgagacaca acgtggcttt cccccccccc cctgcaggtc cgacacgggg
     6361 atggatggcg ttcccgatca tggtcctgct tgcttcgggt ggcatcggaa tgccggcgct
     6421 gcaagcaatg ttgtccaggc aggtggatga ggaacgtcag gggcagctgc aaggctcact
     6481 ggcggcgctc accagcctga cctcgatcgt cggacccctc ctcttcacgg cgatctatgc
     6541 ggcttctata acaacgtgga acgggtgggc atggattgca ggcgctgccc tctacttgct
     6601 ctgcctgccg gcgctgcgtc gcgggctttg gagcggcgca gggcaacgag ccgatcgctg
     6661 atcgtggaaa cgataggcct atgccatgcg ggtcaaggcg acttccggca agctatacgc
     6721 gccctagaat tgtcaatttt aatcctctgt ttatcggcag ttcgtagagc gcgccgtgcg
     6781 tcccgagcga tactgagcga agcaagtgcg tcgagcagtg cccgcttgtt cctgaaatgc
     6841 cagtaaagcg ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt
     6901 gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc
     6961 ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca
     7021 catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc
     7081 gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta
     7141 gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt
     7201 tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc
     7261 aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc
     7321 cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt
     7381 gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac
     7441 cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt
     7501 gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt
     7561 gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc
     7621 aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc
     7681 ggcctgcttg gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt
     7741 ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg
     7801 ttcgagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag
     7861 ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg
     7921 gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc
     7981 ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat
     8041 tcaccctcct tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt
     8101 tgacccgcct ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc
     8161 gtagaccgtc tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac
     8221 cagctccgcg aagtcgctct tcttgatgga gcgcatgggg acgtgcttgg caatcacgcg
     8281 caccccccgg ccgttttagc ggctaaaaaa gtcatggctc tgccctcggg cggaccacgc
     8341 ccatcatgac cttgccaagc tcgtcctgct tctcttcgat cttcgccagc agggcgagga
     8401 tcgtggcatc accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt ttcagcaggc
     8461 cgcccaggcg gcccaggtcg ccattgatgc gggccagctc gcggacgtgc tcatagtcca
     8521 cgacgcccgt gattttgtag ccctggccga cggccagcag gtaggcctac aggctcatgc
     8581 cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag tacaccttga
     8641 taggtgggct gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg
     8701 ttacgccggc ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt
     8761 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
     8821 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
     8881 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
     8941 agggttatgc agcggaaaag atccgtc
//
