LOCUS       40924_24308        8858 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC95, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8858)
  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  JOURNAL   Unpublished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 8858)
  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 8858)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8858)
  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            "Unpublished"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..8858
                     /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(2135..2866)
                     /codon_start=1
                     /gene="icuC"
                     /product="IcuC"
                     /label=icuC
                     /protein_id="ACR44990.1"
                     /translation="MRHTALLAAFLALAAPAAAHDLWINPVGGGVQILYGHPHEPELPN
                     VAKLMSLTAYEPSGAVTLNAKLQTGAAPVLKAAYDGDALVVASYDNGYWVRLSDGSYRN
                     ASKRMLPQADKSLWSVKFAKAVSGPTAPWDKVVGQPLEIVPLEEPAAASGQIRVRVLFE
                     GRPLDGASVVATDGVNFKSEADQARATTDADGVAVVSLRSAGPQVLGVSHRVSPSQTPA
                     LADADSYGATLAFTVTDPKTN"
     gene            complement(2135..2866)
                     /gene="icuC"
                     /label=icuC
     promoter        complement(2894..2924)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     terminator      complement(3029..3056)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3148..3234)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(3416..3432)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3877..4518)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKG"
     CDS             complement(5299..6111)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6786..7931)
                     /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(8203..8574)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(8607..8716)
                     /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 taggctagct gcactcagtt ggtcttcggg tcggttacgg
     2161 tgaaggcgag cgtcgcgccg tagctgtccg catcggccaa cgccggcgtc tgcgaggggc
     2221 tgacgcggtg gctgacgccg agcacctgcg gtccggcact gcgcagcgag acgaccgcga
     2281 cgccgtccgc atccgtcgtg gcgcgggcct gatccgcctc gctcttgaag ttcacgccgt
     2341 cggtggcgac cacgctggcg ccgtcgagcg ggcggccctc gaacagcacg cgcacccgga
     2401 tctggcccga agccgcggcg ggctcctcca gcgggacgat ctccagcggc tggccgacga
     2461 ccttgtccca cggagctgtc gggccggata cggccttggc gaacttcacc gaccagaggc
     2521 tcttgtcggc ctgcggcagc atccgcttgc tggcattgcg gtagctgccg tctgacaagc
     2581 ggacccagta gccgttgtcg taggacgcca cgaccagcgc atcgccgtca taggcggcct
     2641 tgagcacagg tgccgcgccg gtctgcagct tggcgttcag ggtcaccgcg cccgaaggct
     2701 cgtaggcggt caggctcatc agcttggcca cgtttggaag ctccggctcg tggggatgcc
     2761 cgtagaggat ctgcaccccg ccaccgaccg gattgatcca gaggtcgtgg gcggcggccg
     2821 gtgcggcaag ggcgaggaac gcggcgagga gggcggtatg tctcattcgg ggtctctccc
     2881 tggtacctcc acacaacata cgagccggaa gcataaagtg taaagcctgg aattccctgc
     2941 tttcctgatg caaaaacgag gctagtttac cgtatctgtg gggggatggc ttgtagatat
     3001 gacgacagga agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt
     3061 aatttgatgc ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt
     3121 cgcaacgttc aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa
     3181 acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct
     3241 ggcagttccc tactctcgca tggggagacc ccacactacc atcggcgcta cggcgtttca
     3301 cttctgagtt cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaattct
     3361 gttttatcag accgcttctg cgttctgatt taatctgtat caggctgaaa aattcactgg
     3421 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
     3481 cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
     3541 cccaacagtt gcgcagcctg aatggcgaat ggcgcctgat gcggtatttt ctccttacgc
     3601 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     3661 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3721 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3781 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3841 tataggttaa tgtcatgata ataatggttt cttagcaccc tttctcggtc cttcaacgtt
     3901 cctgacaacg agcctccttt tcgccaatcc atcgacaatc accgcgagtc cctgctcgaa
     3961 cgctgcgtcc ggaccggctt cgtcgaaggc gtctatcgcg gcccgcaaca gcggcgagag
     4021 cggagcctgt tcaacggtgc cgccgcgctc gccggcatcg ctgtcgccgg cctgctcctc
     4081 aagcacggcc ccaacagtga agtagctgat tgtcatcagc gcattgacgg cgtccccggc
     4141 cgaaaaaccc gcctcgcaga ggaagcgaag ctgcgcgtcg gccgtttcca tctgcggtgc
     4201 gcccggtcgc gtgccggcat ggatgcgcgc gccatcgcgg taggcgagca gcgcctgcct
     4261 gaagctgcgg gcattcccga tcagaaatga gcgccagtcg tcgtcggctc tcggcaccga
     4321 atgcgtatga ttctccgcca gcatggcttc ggccagtgcg tcgagcagcg cccgcttgtt
     4381 cctgaagtgc cagtaaagcg ccggctgctg aacccccaac cgttccgcca gtttgcgtgt
     4441 cgtcagaccg tctacgccga cctcgttcaa caggtccagg gcggcacgga tcactgtatt
     4501 cggctgcaac tttgtcatga ttgacacttt atcactgata aacataatat gtccaccaac
     4561 ttatcagtga taaagaatcc gcgcgttcaa tcggaccagc ggaggctggt ccggaggcca
     4621 gacgtgaaac ccaacatacc cctgatcgta attctgagca ctgtcgcgct cgacgctgtc
     4681 ggcatcggcc tgattatgcc ggtgctgccg ggcctcctgc gcgatctggt tcactcgaac
     4741 gacgtcaccg cccactatgg cattctgctg gcgctgtatg cgttggtgca atttgcctgc
     4801 gcacctgtgc tgggcgcgct gtcggatcgt ttcgggcggc ggccaatctt gctcgtctcg
     4861 ctggccggcg ccactgtcga ctacgccatc atggcgacag cgcctttcct ttgggttctc
     4921 tatatcgggc ggatcgtggc cggcatcacc ggggcgactg gggcggtagc cggcgcttat
     4981 attgccgatg acctgcaggg gggggggggc gctgaggtct gcctcgtgaa gaaggtgttg
     5041 ctgactcata ccaggcctga atcgccccat catccagcca gaaagtgagg gagccacggt
     5101 tgatgagagc tttgttgtag gtggaccagt tggtgatttt gaacttttgc tttgccacgg
     5161 aacggtctgc gttgtcggga agatgcgtga tctgatcctt caactcagca aaagttcgat
     5221 ttattcaaca aagccgccgt cccgtcaagt cagcgtaatg ctctgccagt gttacaacca
     5281 attaaccaat tctgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat
     5341 atcaggatta tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc
     5401 accgaggcag ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc
     5461 aacatcaata caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc
     5521 accatgagtg acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac
     5581 ttgttcaaca ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt
     5641 attcattcgt gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt
     5701 acaaacagga atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc
     5761 acctgaatca ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt
     5821 gagtaaccat gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa
     5881 ttccgtcagc cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt
     5941 gccatgtttc agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc
     6001 acctgattgc ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt
     6061 ggaatttaat cgcggcctcg agcaagacgt ttcccgttga atatggctca taacacccct
     6121 tgtattactg tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg
     6181 tgcaatgtaa catcagagat tttgagacac aacgtggctt tccccccccc ccctgcaggt
     6241 ccgacacggg gatggatggc gttcccgatc atggtcctgc ttgcttcggg tggcatcgga
     6301 atgccggcgc tgcaagcaat gttgtccagg caggtggatg aggaacgtca ggggcagctg
     6361 caaggctcac tggcggcgct caccagcctg acctcgatcg tcggacccct cctcttcacg
     6421 gcgatctatg cggcttctat aacaacgtgg aacgggtggg catggattgc aggcgctgcc
     6481 ctctacttgc tctgcctgcc ggcgctgcgt cgcgggcttt ggagcggcgc agggcaacga
     6541 gccgatcgct gatcgtggaa acgataggcc tatgccatgc gggtcaaggc gacttccggc
     6601 aagctatacg cgccctagaa ttgtcaattt taatcctctg tttatcggca gttcgtagag
     6661 cgcgccgtgc gtcccgagcg atactgagcg aagcaagtgc gtcgagcagt gcccgcttgt
     6721 tcctgaaatg ccagtaaagc gctggctgct gaacccccag ccggaactga ccccacaagg
     6781 ccctagcgtt tgcaatgcac caggtcatca ttgacccagg cgtgttccac caggccgctg
     6841 cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa
     6901 tccgatccgc acatgaggcg gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag
     6961 ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct tgcggtactt ctcccatatg
     7021 aatttcgtgt agtggtcgcc agcaaacagc acgacgattt cctcgtcgat caggacctgg
     7081 caacgggacg ttttcttgcc acggtccagg acgcggaagc ggtgcagcag cgacaccgat
     7141 tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg
     7201 cattcctcgg ccttcgtgta ataccggcca ttgatcgacc agcccaggtc ctggcaaagc
     7261 tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta ctccaacacc
     7321 tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc
     7381 acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg
     7441 ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc
     7501 ggccacggcg caatatcgaa caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt
     7561 ttcagcaacg cggcctgctt ggcctcgctg acctgttttg ccaggtcctc gccggcggtt
     7621 tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct
     7681 gccgcctcct gttcgagacg acgcgaacgc tccacggcgg ccgatggcgc gggcagggca
     7741 gggggagcca gttgcacgct gtcgcgctcg atcttggccg tagcttgctg gaccatcgag
     7801 ccgacggact ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg
     7861 gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac
     7921 gtccgattca ttcaccctcc ttgcgggatt gccccgactc acgccggggc aatgtgccct
     7981 tattcctgat ttgacccgcc tggtgccttg gtgtccagat aatccacctt atcggcaatg
     8041 aagtcggtcc cgtagaccgt ctggccgtcc ttctcgtact tggtattccg aatcttgccc
     8101 tgcacgaata ccagctccgc gaagtcgctc ttcttgatgg agcgcatggg gacgtgcttg
     8161 gcaatcacgc gcaccccccg gccgttttag cggctaaaaa agtcatggct ctgccctcgg
     8221 gcggaccacg cccatcatga ccttgccaag ctcgtcctgc ttctcttcga tcttcgccag
     8281 cagggcgagg atcgtggcat caccgaaccg cgccgtgcgc gggtcgtcgg tgagccagag
     8341 tttcagcagg ccgcccaggc ggcccaggtc gccattgatg cgggccagct cgcggacgtg
     8401 ctcatagtcc acgacgcccg tgattttgta gccctggccg acggccagca ggtaggccta
     8461 caggctcatg ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt ctggaaggca
     8521 gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag ccatccgctt
     8581 gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat tcccgttgag
     8641 caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg gtgggcctac
     8701 ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta cacgaaccct
     8761 ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa tcgctataat
     8821 gaccccgaag cagggttatg cagcggaaaa gatccgtc
//
