LOCUS       40924_24203        8127 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC41, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8127)
  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  JOURNAL   Unpublished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 8127)
  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 8127)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8127)
  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            "Unpublished"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..8127
                     /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"
     terminator      complement(2298..2325)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(2417..2503)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(2685..2701)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3146..3787)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKG"
     CDS             complement(4568..5380)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6055..7200)
                     /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(7472..7843)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(7876..7985)
                     /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 cttgcatgcc taggctagcg ggcccggatc cgatatctta agatcttccg
     2161 tgcagtttaa gccgaattga tccccgggta ccgagctcga attccctgct ttcctgatgc
     2221 aaaaacgagg ctagtttacc gtatctgtgg ggggatggct tgtagatatg acgacaggaa
     2281 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc
     2341 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc gcaacgttca
     2401 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     2461 aacgaaaggc ccagtctttc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     2521 actctcgcat ggggagaccc cacactacca tcggcgctac ggcgtttcac ttctgagttc
     2581 ggcatggggt caggtgggac caccgcgcta ctgccgccag gcaaattctg ttttatcaga
     2641 ccgcttctgc gttctgattt aatctgtatc aggctgaaaa attcactggc cgtcgtttta
     2701 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     2761 cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg
     2821 cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     2881 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     2941 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     3001 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     3061 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     3121 gtcatgataa taatggtttc ttagcaccct ttctcggtcc ttcaacgttc ctgacaacga
     3181 gcctcctttt cgccaatcca tcgacaatca ccgcgagtcc ctgctcgaac gctgcgtccg
     3241 gaccggcttc gtcgaaggcg tctatcgcgg cccgcaacag cggcgagagc ggagcctgtt
     3301 caacggtgcc gccgcgctcg ccggcatcgc tgtcgccggc ctgctcctca agcacggccc
     3361 caacagtgaa gtagctgatt gtcatcagcg cattgacggc gtccccggcc gaaaaacccg
     3421 cctcgcagag gaagcgaagc tgcgcgtcgg ccgtttccat ctgcggtgcg cccggtcgcg
     3481 tgccggcatg gatgcgcgcg ccatcgcggt aggcgagcag cgcctgcctg aagctgcggg
     3541 cattcccgat cagaaatgag cgccagtcgt cgtcggctct cggcaccgaa tgcgtatgat
     3601 tctccgccag catggcttcg gccagtgcgt cgagcagcgc ccgcttgttc ctgaagtgcc
     3661 agtaaagcgc cggctgctga acccccaacc gttccgccag tttgcgtgtc gtcagaccgt
     3721 ctacgccgac ctcgttcaac aggtccaggg cggcacggat cactgtattc ggctgcaact
     3781 ttgtcatgat tgacacttta tcactgataa acataatatg tccaccaact tatcagtgat
     3841 aaagaatccg cgcgttcaat cggaccagcg gaggctggtc cggaggccag acgtgaaacc
     3901 caacataccc ctgatcgtaa ttctgagcac tgtcgcgctc gacgctgtcg gcatcggcct
     3961 gattatgccg gtgctgccgg gcctcctgcg cgatctggtt cactcgaacg acgtcaccgc
     4021 ccactatggc attctgctgg cgctgtatgc gttggtgcaa tttgcctgcg cacctgtgct
     4081 gggcgcgctg tcggatcgtt tcgggcggcg gccaatcttg ctcgtctcgc tggccggcgc
     4141 cactgtcgac tacgccatca tggcgacagc gcctttcctt tgggttctct atatcgggcg
     4201 gatcgtggcc ggcatcaccg gggcgactgg ggcggtagcc ggcgcttata ttgccgatga
     4261 cctgcagggg ggggggggcg ctgaggtctg cctcgtgaag aaggtgttgc tgactcatac
     4321 caggcctgaa tcgccccatc atccagccag aaagtgaggg agccacggtt gatgagagct
     4381 ttgttgtagg tggaccagtt ggtgattttg aacttttgct ttgccacgga acggtctgcg
     4441 ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa aagttcgatt tattcaacaa
     4501 agccgccgtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     4561 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     4621 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     4681 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     4741 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     4801 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     4861 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     4921 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     4981 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     5041 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     5101 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     5161 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     5221 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     5281 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     5341 gcggcctcga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     5401 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     5461 atcagagatt ttgagacaca acgtggcttt cccccccccc cctgcaggtc cgacacgggg
     5521 atggatggcg ttcccgatca tggtcctgct tgcttcgggt ggcatcggaa tgccggcgct
     5581 gcaagcaatg ttgtccaggc aggtggatga ggaacgtcag gggcagctgc aaggctcact
     5641 ggcggcgctc accagcctga cctcgatcgt cggacccctc ctcttcacgg cgatctatgc
     5701 ggcttctata acaacgtgga acgggtgggc atggattgca ggcgctgccc tctacttgct
     5761 ctgcctgccg gcgctgcgtc gcgggctttg gagcggcgca gggcaacgag ccgatcgctg
     5821 atcgtggaaa cgataggcct atgccatgcg ggtcaaggcg acttccggca agctatacgc
     5881 gccctagaat tgtcaatttt aatcctctgt ttatcggcag ttcgtagagc gcgccgtgcg
     5941 tcccgagcga tactgagcga agcaagtgcg tcgagcagtg cccgcttgtt cctgaaatgc
     6001 cagtaaagcg ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt
     6061 gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc
     6121 ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca
     6181 catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc
     6241 gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta
     6301 gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt
     6361 tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc
     6421 aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc
     6481 cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt
     6541 gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac
     6601 cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt
     6661 gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt
     6721 gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc
     6781 aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc
     6841 ggcctgcttg gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt
     6901 ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg
     6961 ttcgagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag
     7021 ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg
     7081 gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc
     7141 ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat
     7201 tcaccctcct tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt
     7261 tgacccgcct ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc
     7321 gtagaccgtc tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac
     7381 cagctccgcg aagtcgctct tcttgatgga gcgcatgggg acgtgcttgg caatcacgcg
     7441 caccccccgg ccgttttagc ggctaaaaaa gtcatggctc tgccctcggg cggaccacgc
     7501 ccatcatgac cttgccaagc tcgtcctgct tctcttcgat cttcgccagc agggcgagga
     7561 tcgtggcatc accgaaccgc gccgtgcgcg ggtcgtcggt gagccagagt ttcagcaggc
     7621 cgcccaggcg gcccaggtcg ccattgatgc gggccagctc gcggacgtgc tcatagtcca
     7681 cgacgcccgt gattttgtag ccctggccga cggccagcag gtaggcctac aggctcatgc
     7741 cggccgccgc cgccttttcc tcaatcgctc ttcgttcgtc tggaaggcag tacaccttga
     7801 taggtgggct gcccttcctg gttggcttgg tttcatcagc catccgcttg ccctcatctg
     7861 ttacgccggc ggtagccggc cagcctcgca gagcaggatt cccgttgagc accgccaggt
     7921 gcgaataagg gacagtgaag aaggaacacc cgctcgcggg tgggcctact tcacctatcc
     7981 tgcccggctg acgccgttgg atacaccaag gaaagtctac acgaaccctt tggcaaaatc
     8041 ctgtatatcg tgcgaaaaag gatggatata ccgaaaaaat cgctataatg accccgaagc
     8101 agggttatgc agcggaaaag atccgtc
//
