LOCUS       40924_24283        8865 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC85, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8865)
  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  JOURNAL   Unpublished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 8865)
  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 8865)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8865)
  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            "Unpublished"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..8865
                     /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="ACR44975.1"
                     /translation="MRHTALLAAFLALAAPAAAHDLWINPVGGGVQILYGHPHEPELPN
                     VAKLMSLTAYEPSGAVTLNAKLQTGAAPVLKAAYDGDALVVASYDNGYWVRLSDGSYRN
                     ASKRMLPQADKSLWSVKFAKAVSGPTAPWDKVVGQPLEIVPLEEPAAASGQIRVRVLFE
                     GRPLDGASVVATDGVNFKSEADQARATTDADGVAVVSLRSAGPQVLGVSHRVSPSQTPA
                     LADADSYGATLAFTVTDPKTN"
     gene            complement(2135..2866)
                     /gene="icuC"
                     /label=icuC
     promoter        complement(2894..2922)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     terminator      complement(3036..3063)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(3155..3241)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(3423..3439)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3884..4525)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKG"
     CDS             complement(5306..6118)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6793..7938)
                     /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(8210..8581)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(8614..8723)
                     /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 acacattata cgagccgatg attaattgtc aacagctcat ttcagagaat
     2941 tccctgcttt cctgatgcaa aaacgaggct agtttaccgt atctgtgggg ggatggcttg
     3001 tagatatgac gacaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
     3061 tctgcttaat ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
     3121 gttgcttcgc aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
     3181 ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt cgttttattt
     3241 gatgcctggc agttccctac tctcgcatgg ggagacccca cactaccatc ggcgctacgg
     3301 cgtttcactt ctgagttcgg catggggtca ggtgggacca ccgcgctact gccgccaggc
     3361 aaattctgtt ttatcagacc gcttctgcgt tctgatttaa tctgtatcag gctgaaaaat
     3421 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     3481 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     3541 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     3601 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     3661 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     3721 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     3781 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     3841 ctatttttat aggttaatgt catgataata atggtttctt agcacccttt ctcggtcctt
     3901 caacgttcct gacaacgagc ctccttttcg ccaatccatc gacaatcacc gcgagtccct
     3961 gctcgaacgc tgcgtccgga ccggcttcgt cgaaggcgtc tatcgcggcc cgcaacagcg
     4021 gcgagagcgg agcctgttca acggtgccgc cgcgctcgcc ggcatcgctg tcgccggcct
     4081 gctcctcaag cacggcccca acagtgaagt agctgattgt catcagcgca ttgacggcgt
     4141 ccccggccga aaaacccgcc tcgcagagga agcgaagctg cgcgtcggcc gtttccatct
     4201 gcggtgcgcc cggtcgcgtg ccggcatgga tgcgcgcgcc atcgcggtag gcgagcagcg
     4261 cctgcctgaa gctgcgggca ttcccgatca gaaatgagcg ccagtcgtcg tcggctctcg
     4321 gcaccgaatg cgtatgattc tccgccagca tggcttcggc cagtgcgtcg agcagcgccc
     4381 gcttgttcct gaagtgccag taaagcgccg gctgctgaac ccccaaccgt tccgccagtt
     4441 tgcgtgtcgt cagaccgtct acgccgacct cgttcaacag gtccagggcg gcacggatca
     4501 ctgtattcgg ctgcaacttt gtcatgattg acactttatc actgataaac ataatatgtc
     4561 caccaactta tcagtgataa agaatccgcg cgttcaatcg gaccagcgga ggctggtccg
     4621 gaggccagac gtgaaaccca acatacccct gatcgtaatt ctgagcactg tcgcgctcga
     4681 cgctgtcggc atcggcctga ttatgccggt gctgccgggc ctcctgcgcg atctggttca
     4741 ctcgaacgac gtcaccgccc actatggcat tctgctggcg ctgtatgcgt tggtgcaatt
     4801 tgcctgcgca cctgtgctgg gcgcgctgtc ggatcgtttc gggcggcggc caatcttgct
     4861 cgtctcgctg gccggcgcca ctgtcgacta cgccatcatg gcgacagcgc ctttcctttg
     4921 ggttctctat atcgggcgga tcgtggccgg catcaccggg gcgactgggg cggtagccgg
     4981 cgcttatatt gccgatgacc tgcagggggg ggggggcgct gaggtctgcc tcgtgaagaa
     5041 ggtgttgctg actcatacca ggcctgaatc gccccatcat ccagccagaa agtgagggag
     5101 ccacggttga tgagagcttt gttgtaggtg gaccagttgg tgattttgaa cttttgcttt
     5161 gccacggaac ggtctgcgtt gtcgggaaga tgcgtgatct gatccttcaa ctcagcaaaa
     5221 gttcgattta ttcaacaaag ccgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt
     5281 acaaccaatt aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt
     5341 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
     5401 aaaactcacc gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga
     5461 ctcgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
     5521 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caaaagctta tgcatttctt
     5581 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
     5641 aaccgttatt cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag
     5701 gacaattaca aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa
     5761 tattttcacc tgaatcagga tattcttcta atacctggaa tgctgttttc ccggggatcg
     5821 cagtggtgag taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag
     5881 gcataaattc cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc
     5941 tacctttgcc atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcgataga
     6001 ttgtcgcacc tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat
     6061 ccatgttgga atttaatcgc ggcctcgagc aagacgtttc ccgttgaata tggctcataa
     6121 caccccttgt attactgttt atgtaagcag acagttttat tgttcatgat gatatatttt
     6181 tatcttgtgc aatgtaacat cagagatttt gagacacaac gtggctttcc cccccccccc
     6241 tgcaggtccg acacggggat ggatggcgtt cccgatcatg gtcctgcttg cttcgggtgg
     6301 catcggaatg ccggcgctgc aagcaatgtt gtccaggcag gtggatgagg aacgtcaggg
     6361 gcagctgcaa ggctcactgg cggcgctcac cagcctgacc tcgatcgtcg gacccctcct
     6421 cttcacggcg atctatgcgg cttctataac aacgtggaac gggtgggcat ggattgcagg
     6481 cgctgccctc tacttgctct gcctgccggc gctgcgtcgc gggctttgga gcggcgcagg
     6541 gcaacgagcc gatcgctgat cgtggaaacg ataggcctat gccatgcggg tcaaggcgac
     6601 ttccggcaag ctatacgcgc cctagaattg tcaattttaa tcctctgttt atcggcagtt
     6661 cgtagagcgc gccgtgcgtc ccgagcgata ctgagcgaag caagtgcgtc gagcagtgcc
     6721 cgcttgttcc tgaaatgcca gtaaagcgct ggctgctgaa cccccagccg gaactgaccc
     6781 cacaaggccc tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag
     6841 gccgctgcct cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg
     6901 ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg
     6961 gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc
     7021 ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag
     7081 gacctggcaa cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga
     7141 caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg
     7201 cgacaggcat tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg
     7261 gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc
     7321 caacacctgc tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt
     7381 gatcttcacg tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat
     7441 tttcttgttg cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat
     7501 cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt
     7561 cgtgtgtttc agcaacgcgg cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc
     7621 ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc
     7681 caaacctgcc gcctcctgtt cgagacgacg cgaacgctcc acggcggccg atggcgcggg
     7741 cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac
     7801 catcgagccg acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat
     7861 ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg
     7921 gtcaaacgtc cgattcattc accctccttg cgggattgcc ccgactcacg ccggggcaat
     7981 gtgcccttat tcctgatttg acccgcctgg tgccttggtg tccagataat ccaccttatc
     8041 ggcaatgaag tcggtcccgt agaccgtctg gccgtccttc tcgtacttgg tattccgaat
     8101 cttgccctgc acgaatacca gctccgcgaa gtcgctcttc ttgatggagc gcatggggac
     8161 gtgcttggca atcacgcgca ccccccggcc gttttagcgg ctaaaaaagt catggctctg
     8221 ccctcgggcg gaccacgccc atcatgacct tgccaagctc gtcctgcttc tcttcgatct
     8281 tcgccagcag ggcgaggatc gtggcatcac cgaaccgcgc cgtgcgcggg tcgtcggtga
     8341 gccagagttt cagcaggccg cccaggcggc ccaggtcgcc attgatgcgg gccagctcgc
     8401 ggacgtgctc atagtccacg acgcccgtga ttttgtagcc ctggccgacg gccagcaggt
     8461 aggcctacag gctcatgccg gccgccgccg ccttttcctc aatcgctctt cgttcgtctg
     8521 gaaggcagta caccttgata ggtgggctgc ccttcctggt tggcttggtt tcatcagcca
     8581 tccgcttgcc ctcatctgtt acgccggcgg tagccggcca gcctcgcaga gcaggattcc
     8641 cgttgagcac cgccaggtgc gaataaggga cagtgaagaa ggaacacccg ctcgcgggtg
     8701 ggcctacttc acctatcctg cccggctgac gccgttggat acaccaagga aagtctacac
     8761 gaaccctttg gcaaaatcct gtatatcgtg cgaaaaagga tggatatacc gaaaaaatcg
     8821 ctataatgac cccgaagcag ggttatgcag cggaaaagat ccgtc
//
