LOCUS       Exported               10253 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pHC69, complete sequence.
ACCESSION   FJ389172
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10253)
  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  JOURNAL   Unpublished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 10253)
  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 10253)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10253
                     /organism="Cloning vector pHC69"
                     /mol_type="other DNA"
                     /db_xref="taxon:646118"
     rep_origin      4..712
                     /label=oriV
                     /note="oriV"
     rep_origin      11..642
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      1180..1735
                     /label=colE1
                     /note="colE1"
     protein_bind    1981..2002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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(2140..4254)
                     /codon_start=1
                     /gene="icuA"
                     /product="IcuA"
                     /label=icuA
                     /protein_id="ACR44965.1"
                     /translation="MSPLLTRLLASASALGVSTLPSHALDPTAVELEELQVTAQGGSAN
                     SVAAVAGPAANLKTQISPSVLALPGNSTVIDAAEIARLPVQSYGDLFRPVAGFDVSNYG
                     QGGVGYGIALRGFSDAEHGRDVAVFIDGVPINEVSSIHTPNYVDLNPLIPETIRSISIV
                     RGPFSVEAGDANLGGSIFIETKRAEPFSTVGASGGSFGTWRGLATFSQIGPGIQPFLAY
                     EGYGTGGYRENGDLGRYNAFNKVTIPLDLGSSLSLRVQMYGTGFGQPGYLERDRVRAGL
                     VSPRAATNITDGGGKTLQTFVANYVNGLPDDELSTLLYASHDVFSRFSDFGAGQRGQVE
                     ERETVGGRIRKVWTTSLFDVLPTQVLVGGDWRSDALDVNTGPSVARTFTARTLDLDLSQ
                     HNLAAYAQVQVKPAPWLKLTGGARFDQFFYDVRNNLDPALSPKVSPNVVSPKGGAALTP
                     FDWLELFANYGQGFRSPSASSELTNNPRLTPLTIESTEGGGRLVFDRFSLLGTVFFTDI
                     ANEVFQPAPGLPVQNLGKSRRDGFEIEAKVLAYREGLDRVALFANYSGVSARLLNGAPS
                     IFIPNVPAYLATVGAEFDVAAPAALLPGGRLTGAAYLSYIGAKPLTEDGVLHSSPYPRV
                     SGRIAYVHQSGWTTYGQATWYPGDRTSESIFNFGSSVSASAAEIVTSPQPRLTLLAGIT
                     SSFATGAPLR"
     gene            complement(2140..4254)
                     /gene="icuA"
                     /label=icuA
     promoter        complement(4282..4310)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      4342..4805
                     /regulatory_class="terminator"
                     /note="rrnB"
     terminator      4424..4451
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      4543..4629
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4811..4827)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(6691..7506)
                     /codon_start=1
                     /gene="aph"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=aph
                     /note="Aph; kanamycin-resistance protein"
                     /protein_id="ACR44966.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(6691..7506)
                     /gene="aph"
                     /label=aph
     CDS             complement(8178..9326)
                     /codon_start=1
                     /gene="trfA"
                     /product="plasmid replication protein TrfA"
                     /label=trfA
                     /protein_id="ACR44967.1"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     gene            complement(8178..9326)
                     /gene="trfA"
                     /label=trfA
     oriT            9508..10246
                     /label=origin of transfer
                     /note="origin of transfer"
     CDS             complement(9598..9969)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            10002..10111
                     /note="incP origin of transfer"