LOCUS       40924_24268        8863 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC70, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8863)
  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  JOURNAL   Unpublished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 8863)
  AUTHORS   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 8863)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8863)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     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"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8863
                     /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(2133..2864)
                     /codon_start=1
                     /gene="icuB"
                     /product="IcuB"
                     /label=icuB
                     /protein_id="ACR44968.1"
                     /translation="MRRTVILTALLAFTAPAAAHDLWLDPAGNGVQILYGHPHEPELPS
                     AGKLMSLTAYEPSGAVALNAKLQTGAMPALKAAHQGDALFAAAYDNGYWVRLSDGGYRN
                     ASKRMLPQADKSLWSVKFAKAVSGPTAPWDKIVGQPLEIVPLEEPAAASGQIRVRVLFE
                     GRPLDGASVVATDGVNFKSEADQARATTDADGVAVVSLRSAGPQVLGVAHRVSPSQTPA
                     LADADSYGATLAFTVTDPKSN"
     gene            complement(2133..2864)
                     /gene="icuB"
                     /label=icuB
     promoter        complement(2892..2920)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     terminator      complement(3034..3061)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3153..3239)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(3421..3437)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3882..4523)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKG"
     CDS             complement(5304..6116)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6791..7936)
                     /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(8208..8579)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(8612..8721)
                     /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 taggctagcc gctcagttgg acttcgggtc ggtcacggtg
     2161 aaggcgagcg tcgcgccgta gctgtccgca tcggccagcg ccggcgtctg cgaggggctc
     2221 acgcggtgcg ccacgccgag cacctgggga ccggcactgc gcagcgagac gaccgccacg
     2281 ccgtccgcat ccgtcgtggc gcgggcctga tccgcctcgc tcttgaagtt cacgccgtcg
     2341 gtggcgacca cgctggcgcc gtcgagcgga cggccctcga acagcacccg cacccggatc
     2401 tggcccgaag ccgcggccgg ctcctccagc gggacgatct ccagcggttg gccgacgatc
     2461 ttgtcccacg gggccgtcgg gccggatacg gccttggcga acttcaccga ccagaggctc
     2521 ttgtcggcct gcggcagcat ccgcttgctg gcgttgcggt agccgccgtc ggacaagcgc
     2581 acccagtagc cgttgtcgta ggccgcggcg aacagcgcgt cgccctgatg cgcggctttg
     2641 agcgcgggca ttgcgccggt ctgcagcttg gcattcaggg ccaccgcgcc ggacggctcg
     2701 taggcggtca ggctcatcag cttgccagca ctcggcagtt ccggctcgtg gggatgcccg
     2761 tagaggatct gcacgccgtt gccggccggg tcgagccaga ggtcgtgggc ggcggccggt
     2821 gcggtgaagg cgagcagcgc ggtgaggatg acggttcgtc tcattcgggg tctctccctg
     2881 gtacctccac acattatacg agccgatgat taattgtcaa cagctcattt cagagaattc
     2941 cctgctttcc tgatgcaaaa acgaggctag tttaccgtat ctgtgggggg atggcttgta
     3001 gatatgacga caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     3061 tgcttaattt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     3121 tgcttcgcaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     3181 gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg ttttatttga
     3241 tgcctggcag ttccctactc tcgcatgggg agaccccaca ctaccatcgg cgctacggcg
     3301 tttcacttct gagttcggca tggggtcagg tgggaccacc gcgctactgc cgccaggcaa
     3361 attctgtttt atcagaccgc ttctgcgttc tgatttaatc tgtatcaggc tgaaaaattc
     3421 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     3481 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     3541 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt attttctcct
     3601 tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa tctgctctga
     3661 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3721 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3781 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3841 atttttatag gttaatgtca tgataataat ggtttcttag caccctttct cggtccttca
     3901 acgttcctga caacgagcct ccttttcgcc aatccatcga caatcaccgc gagtccctgc
     3961 tcgaacgctg cgtccggacc ggcttcgtcg aaggcgtcta tcgcggcccg caacagcggc
     4021 gagagcggag cctgttcaac ggtgccgccg cgctcgccgg catcgctgtc gccggcctgc
     4081 tcctcaagca cggccccaac agtgaagtag ctgattgtca tcagcgcatt gacggcgtcc
     4141 ccggccgaaa aacccgcctc gcagaggaag cgaagctgcg cgtcggccgt ttccatctgc
     4201 ggtgcgcccg gtcgcgtgcc ggcatggatg cgcgcgccat cgcggtaggc gagcagcgcc
     4261 tgcctgaagc tgcgggcatt cccgatcaga aatgagcgcc agtcgtcgtc ggctctcggc
     4321 accgaatgcg tatgattctc cgccagcatg gcttcggcca gtgcgtcgag cagcgcccgc
     4381 ttgttcctga agtgccagta aagcgccggc tgctgaaccc ccaaccgttc cgccagtttg
     4441 cgtgtcgtca gaccgtctac gccgacctcg ttcaacaggt ccagggcggc acggatcact
     4501 gtattcggct gcaactttgt catgattgac actttatcac tgataaacat aatatgtcca
     4561 ccaacttatc agtgataaag aatccgcgcg ttcaatcgga ccagcggagg ctggtccgga
     4621 ggccagacgt gaaacccaac atacccctga tcgtaattct gagcactgtc gcgctcgacg
     4681 ctgtcggcat cggcctgatt atgccggtgc tgccgggcct cctgcgcgat ctggttcact
     4741 cgaacgacgt caccgcccac tatggcattc tgctggcgct gtatgcgttg gtgcaatttg
     4801 cctgcgcacc tgtgctgggc gcgctgtcgg atcgtttcgg gcggcggcca atcttgctcg
     4861 tctcgctggc cggcgccact gtcgactacg ccatcatggc gacagcgcct ttcctttggg
     4921 ttctctatat cgggcggatc gtggccggca tcaccggggc gactggggcg gtagccggcg
     4981 cttatattgc cgatgacctg cagggggggg ggggcgctga ggtctgcctc gtgaagaagg
     5041 tgttgctgac tcataccagg cctgaatcgc cccatcatcc agccagaaag tgagggagcc
     5101 acggttgatg agagctttgt tgtaggtgga ccagttggtg attttgaact tttgctttgc
     5161 cacggaacgg tctgcgttgt cgggaagatg cgtgatctga tccttcaact cagcaaaagt
     5221 tcgatttatt caacaaagcc gccgtcccgt caagtcagcg taatgctctg ccagtgttac
     5281 aaccaattaa ccaattctga ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta
     5341 ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa
     5401 aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact
     5461 cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag
     5521 aaatcaccat gagtgacgac tgaatccggt gagaatggca aaagcttatg catttctttc
     5581 cagacttgtt caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa
     5641 ccgttattca ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga
     5701 caattacaaa caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata
     5761 ttttcacctg aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca
     5821 gtggtgagta accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc
     5881 ataaattccg tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta
     5941 cctttgccat gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt
     6001 gtcgcacctg attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc
     6061 atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcataaca
     6121 ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta
     6181 tcttgtgcaa tgtaacatca gagattttga gacacaacgt ggctttcccc cccccccctg
     6241 caggtccgac acggggatgg atggcgttcc cgatcatggt cctgcttgct tcgggtggca
     6301 tcggaatgcc ggcgctgcaa gcaatgttgt ccaggcaggt ggatgaggaa cgtcaggggc
     6361 agctgcaagg ctcactggcg gcgctcacca gcctgacctc gatcgtcgga cccctcctct
     6421 tcacggcgat ctatgcggct tctataacaa cgtggaacgg gtgggcatgg attgcaggcg
     6481 ctgccctcta cttgctctgc ctgccggcgc tgcgtcgcgg gctttggagc ggcgcagggc
     6541 aacgagccga tcgctgatcg tggaaacgat aggcctatgc catgcgggtc aaggcgactt
     6601 ccggcaagct atacgcgccc tagaattgtc aattttaatc ctctgtttat cggcagttcg
     6661 tagagcgcgc cgtgcgtccc gagcgatact gagcgaagca agtgcgtcga gcagtgcccg
     6721 cttgttcctg aaatgccagt aaagcgctgg ctgctgaacc cccagccgga actgacccca
     6781 caaggcccta gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc
     6841 cgctgcctcg caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg
     6901 tggaatccga tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt
     6961 gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc
     7021 atatgaattt cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga
     7081 cctggcaacg ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca
     7141 ccgattccag gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg
     7201 acaggcattc ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc
     7261 aaagctcgta gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca
     7321 acacctgctg ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga
     7381 tcttcacgtc cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt
     7441 tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg
     7501 tgtccggcca cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg
     7561 tgtgtttcag caacgcggcc tgcttggcct cgctgacctg ttttgccagg tcctcgccgg
     7621 cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca
     7681 aacctgccgc ctcctgttcg agacgacgcg aacgctccac ggcggccgat ggcgcgggca
     7741 gggcaggggg agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacca
     7801 tcgagccgac ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg
     7861 tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt
     7921 caaacgtccg attcattcac cctccttgcg ggattgcccc gactcacgcc ggggcaatgt
     7981 gcccttattc ctgatttgac ccgcctggtg ccttggtgtc cagataatcc accttatcgg
     8041 caatgaagtc ggtcccgtag accgtctggc cgtccttctc gtacttggta ttccgaatct
     8101 tgccctgcac gaataccagc tccgcgaagt cgctcttctt gatggagcgc atggggacgt
     8161 gcttggcaat cacgcgcacc ccccggccgt tttagcggct aaaaaagtca tggctctgcc
     8221 ctcgggcgga ccacgcccat catgaccttg ccaagctcgt cctgcttctc ttcgatcttc
     8281 gccagcaggg cgaggatcgt ggcatcaccg aaccgcgccg tgcgcgggtc gtcggtgagc
     8341 cagagtttca gcaggccgcc caggcggccc aggtcgccat tgatgcgggc cagctcgcgg
     8401 acgtgctcat agtccacgac gcccgtgatt ttgtagccct ggccgacggc cagcaggtag
     8461 gcctacaggc tcatgccggc cgccgccgcc ttttcctcaa tcgctcttcg ttcgtctgga
     8521 aggcagtaca ccttgatagg tgggctgccc ttcctggttg gcttggtttc atcagccatc
     8581 cgcttgccct catctgttac gccggcggta gccggccagc ctcgcagagc aggattcccg
     8641 ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg aacacccgct cgcgggtggg
     8701 cctacttcac ctatcctgcc cggctgacgc cgttggatac accaaggaaa gtctacacga
     8761 accctttggc aaaatcctgt atatcgtgcg aaaaaggatg gatataccga aaaaatcgct
     8821 ataatgaccc cgaagcaggg ttatgcagcg gaaaagatcc gtc
//
