LOCUS       40924_24288        8163 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC91, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8163)
  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  JOURNAL   Unpublished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 8163)
  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 8163)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8163)
  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            "Unpublished"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..8163
                     /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"
     promoter        complement(2199..2229)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     terminator      complement(2334..2361)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(2453..2539)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     complement(2721..2737)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3182..3823)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKG"
     CDS             complement(4604..5416)
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(6091..7236)
                     /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(7508..7879)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(7912..8021)
                     /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 cctccacaca acatacgagc cggaagcata
     2221 aagtgtaaag cctggaattc cctgctttcc tgatgcaaaa acgaggctag tttaccgtat
     2281 ctgtgggggg atggcttgta gatatgacga caggaagagt ttgtagaaac gcaaaaaggc
     2341 catccgtcag gatggccttc tgcttaattt gatgcctggc agtttatggc gggcgtcctg
     2401 cccgccaccc tccgggccgt tgcttcgcaa cgttcaaatc cgctcccggc ggatttgtcc
     2461 tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact
     2521 gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcatgggg agaccccaca
     2581 ctaccatcgg cgctacggcg tttcacttct gagttcggca tggggtcagg tgggaccacc
     2641 gcgctactgc cgccaggcaa attctgtttt atcagaccgc ttctgcgttc tgatttaatc
     2701 tgtatcaggc tgaaaaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     2761 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     2821 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc
     2881 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
     2941 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     3001 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3061 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     3121 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     3181 caccctttct cggtccttca acgttcctga caacgagcct ccttttcgcc aatccatcga
     3241 caatcaccgc gagtccctgc tcgaacgctg cgtccggacc ggcttcgtcg aaggcgtcta
     3301 tcgcggcccg caacagcggc gagagcggag cctgttcaac ggtgccgccg cgctcgccgg
     3361 catcgctgtc gccggcctgc tcctcaagca cggccccaac agtgaagtag ctgattgtca
     3421 tcagcgcatt gacggcgtcc ccggccgaaa aacccgcctc gcagaggaag cgaagctgcg
     3481 cgtcggccgt ttccatctgc ggtgcgcccg gtcgcgtgcc ggcatggatg cgcgcgccat
     3541 cgcggtaggc gagcagcgcc tgcctgaagc tgcgggcatt cccgatcaga aatgagcgcc
     3601 agtcgtcgtc ggctctcggc accgaatgcg tatgattctc cgccagcatg gcttcggcca
     3661 gtgcgtcgag cagcgcccgc ttgttcctga agtgccagta aagcgccggc tgctgaaccc
     3721 ccaaccgttc cgccagtttg cgtgtcgtca gaccgtctac gccgacctcg ttcaacaggt
     3781 ccagggcggc acggatcact gtattcggct gcaactttgt catgattgac actttatcac
     3841 tgataaacat aatatgtcca ccaacttatc agtgataaag aatccgcgcg ttcaatcgga
     3901 ccagcggagg ctggtccgga ggccagacgt gaaacccaac atacccctga tcgtaattct
     3961 gagcactgtc gcgctcgacg ctgtcggcat cggcctgatt atgccggtgc tgccgggcct
     4021 cctgcgcgat ctggttcact cgaacgacgt caccgcccac tatggcattc tgctggcgct
     4081 gtatgcgttg gtgcaatttg cctgcgcacc tgtgctgggc gcgctgtcgg atcgtttcgg
     4141 gcggcggcca atcttgctcg tctcgctggc cggcgccact gtcgactacg ccatcatggc
     4201 gacagcgcct ttcctttggg ttctctatat cgggcggatc gtggccggca tcaccggggc
     4261 gactggggcg gtagccggcg cttatattgc cgatgacctg cagggggggg ggggcgctga
     4321 ggtctgcctc gtgaagaagg tgttgctgac tcataccagg cctgaatcgc cccatcatcc
     4381 agccagaaag tgagggagcc acggttgatg agagctttgt tgtaggtgga ccagttggtg
     4441 attttgaact tttgctttgc cacggaacgg tctgcgttgt cgggaagatg cgtgatctga
     4501 tccttcaact cagcaaaagt tcgatttatt caacaaagcc gccgtcccgt caagtcagcg
     4561 taatgctctg ccagtgttac aaccaattaa ccaattctga ttagaaaaac tcatcgagca
     4621 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     4681 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     4741 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     4801 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     4861 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     4921 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     4981 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     5041 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     5101 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     5161 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     5221 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     5281 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     5341 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc
     5401 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     5461 ttcatgatga tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaacgt
     5521 ggctttcccc cccccccctg caggtccgac acggggatgg atggcgttcc cgatcatggt
     5581 cctgcttgct tcgggtggca tcggaatgcc ggcgctgcaa gcaatgttgt ccaggcaggt
     5641 ggatgaggaa cgtcaggggc agctgcaagg ctcactggcg gcgctcacca gcctgacctc
     5701 gatcgtcgga cccctcctct tcacggcgat ctatgcggct tctataacaa cgtggaacgg
     5761 gtgggcatgg attgcaggcg ctgccctcta cttgctctgc ctgccggcgc tgcgtcgcgg
     5821 gctttggagc ggcgcagggc aacgagccga tcgctgatcg tggaaacgat aggcctatgc
     5881 catgcgggtc aaggcgactt ccggcaagct atacgcgccc tagaattgtc aattttaatc
     5941 ctctgtttat cggcagttcg tagagcgcgc cgtgcgtccc gagcgatact gagcgaagca
     6001 agtgcgtcga gcagtgcccg cttgttcctg aaatgccagt aaagcgctgg ctgctgaacc
     6061 cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt catcattgac
     6121 ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc cgacctgctc
     6181 gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg tttccagctt
     6241 gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga
     6301 cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa acagcacgac
     6361 gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt ccaggacgcg
     6421 gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg tgaagcccat
     6481 cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc ggccattgat
     6541 cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct cgccgatagg
     6601 ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt cggcccgcag
     6661 ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga ccttgttttg
     6721 cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc
     6781 gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg aaagctgcat
     6841 ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct cgctgacctg
     6901 ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc
     6961 gatggtcatc gacttcgcca aacctgccgc ctcctgttcg agacgacgcg aacgctccac
     7021 ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc gctcgatctt
     7081 ggccgtagct tgctggacca tcgagccgac ggactggaag gtttcgcggg gcgcacgcat
     7141 gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc
     7201 ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg ggattgcccc
     7261 gactcacgcc ggggcaatgt gcccttattc ctgatttgac ccgcctggtg ccttggtgtc
     7321 cagataatcc accttatcgg caatgaagtc ggtcccgtag accgtctggc cgtccttctc
     7381 gtacttggta ttccgaatct tgccctgcac gaataccagc tccgcgaagt cgctcttctt
     7441 gatggagcgc atggggacgt gcttggcaat cacgcgcacc ccccggccgt tttagcggct
     7501 aaaaaagtca tggctctgcc ctcgggcgga ccacgcccat catgaccttg ccaagctcgt
     7561 cctgcttctc ttcgatcttc gccagcaggg cgaggatcgt ggcatcaccg aaccgcgccg
     7621 tgcgcgggtc gtcggtgagc cagagtttca gcaggccgcc caggcggccc aggtcgccat
     7681 tgatgcgggc cagctcgcgg acgtgctcat agtccacgac gcccgtgatt ttgtagccct
     7741 ggccgacggc cagcaggtag gcctacaggc tcatgccggc cgccgccgcc ttttcctcaa
     7801 tcgctcttcg ttcgtctgga aggcagtaca ccttgatagg tgggctgccc ttcctggttg
     7861 gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc
     7921 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
     7981 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
     8041 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
     8101 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaagatcc
     8161 gtc
//
