LOCUS       Exported                3990 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pMCI002, complete sequence.
ACCESSION   JN788332
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3990)
  AUTHORS   Uhlich GA, Chen CY.
  TITLE     A cloning vector for creation of Escherichia colilacZ translational 
            fusions and generation of linear template for chromosomal 
            integration
  JOURNAL   Plasmid 67 (3), 259-263 (2012)
  PUBMED    22197962
REFERENCE   2  (bases 1 to 3990)
  AUTHORS   Chen C-Y., Uhlich GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-SEP-2011) Molecular Characterization of Foodborne 
            Pathogens RU, US Department of Agriculture, 600 E. Mermaid Lane, 
            Wyndmoor, PA 19038, USA
REFERENCE   3  (bases 1 to 3990)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3990
                     /organism="Cloning vector pMCI002"
                     /mol_type="other DNA"
                     /label=propagated in EC100Dpir+
                     /note="propagated in EC100Dpir+"
                     /db_xref="taxon:1145073"
     primer_bind     1..21
                     /label=LacIF primer binding site
                     /note="LacIF primer binding site"
     primer_bind     51..76
                     /note="SpecF primer binding site; primer with additional 
                     sequence at the 5 end AAGGAATT"
     CDS             228..1010
                     /codon_start=1
                     /gene="ANT(9)"
                     /product="spectinomycin 3-adenylyltransferase"
                     /label=ANT(9)
                     /note="spectinomycin resistance"
                     /protein_id="AFA52509.1"
                     /translation="MSNLINGKIPNQAIQTLKIVKDLFGSSIVGVYLFGSAVNGGLRIN
                     SDVDVLVVVNHSLPQLTRKKLTERLMTISGKIGNTDSVRPLEVTVINRSEVVPWQYPPK
                     REFIYGEWLRGEFENGQIQEPSYDPDLAIVLAQARKNSISLFGPDSSSILVSVPLTDIR
                     RAIKDSLPELIEGIKGDERNVILTLARMWQTVTTGEITSKDVAAEWAIPLLPKEHVTLL
                     DIARKGYRGECDDKWEGLYSKVKALVKYMKNSIETSLN"
     gene            228..1010
                     /gene="ANT(9)"
                     /label=ANT(9)
     primer_bind     complement(1257..1294)
                     /label=SpecR primer binding site
                     /note="SpecR primer binding site"
     misc_feature    1279..1302
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             1298..1408
                     /codon_start=1
                     /product="lacZ"
                     /label=lacZ
                     /note="lacZ reading frame for translational fusion"
                     /protein_id="AFA52512.1"
                     /translation="DPVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR"
     primer_bind     complement(1346..1367)
                     /label=LacZR primer binding site
                     /note="LacZR primer binding site"
     primer_bind     complement(1389..1409)
                     /label=LacZR2 primer binding site
                     /note="LacZR2 primer binding site"
     misc_feature    1410..3990
                     /label=from plasmid pRE112, EcoRI filled-in
                     /note="from plasmid pRE112, EcoRI filled-in"
     rep_origin      1415..1806
                     /note="R6K origin; similar to GenBank Accession number 
                     V00623"
     rep_origin      1418..1806
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    1836..3563
                     /note="mob RP4, similar to RP4 plasmid sequence (GenBank 
                     Accession Number X54459)"
     CDS             complement(1836..2393)
                     /codon_start=1
                     /gene="traI"
                     /product="TraI"
                     /label=traI
                     /note="TraI DNA relaxase"
                     /protein_id="AFA52510.1"
                     /translation="MIAKHVPMRSIKKSDFAELVKYITDEQGKTERLGHVRVTNCEANT
                     LPAVMAEVMATQHGNTRSEADKTYHLLVSFRAGEKPDAETLRAIEDRICAGLGFAEHQR
                     VSAVHHDTDNLHIHIAINKIHPTRNTIHEPYRAYRALADLCATLERDYGLERDNHETRQ
                     RVSENRANDMERHAGVESLVGWI"
     gene            complement(1836..2393)
                     /gene="traI"
                     /label=traI
     CDS             complement(2428..2799)
                     /codon_start=1
                     /gene="traJ"
                     /product="TraJ"
                     /label=traJ
                     /note="oriT recognition protein"
                     /protein_id="AFA52511.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            complement(2428..2799)
                     /gene="traJ"
                     /label=traJ
     oriT            complement(2832..2943)
                     /direction=LEFT
                     /label=RP4 origin of transfer
                     /note="RP4 origin of transfer"
     oriT            2832..2941
                     /note="incP origin of transfer"
     CDS             3167..3563
                     /codon_start=1
                     /gene="traK"
                     /product="TraK"
                     /label=traK
                     /note="oriT binding protein"
                     /protein_id="AFA52508.1"
                     /translation="MPKSYTDELAEWVESRAAKKRRRDEAAVAFLAVRADVEAALASGY
                     ALVTIWEHMRETGKVKFSYETFRSHARRHIKAKPADVPAPQAKAAEPAPAPKTPEPRRP
                     KQGGKAEKPAPAAAPTGFTFNPTPDKKD"
     gene            3167..3563
                     /gene="traK"
                     /label=traK
     promoter        3670..3772
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"