pKIKOlacZKm vector (V005239)

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LOCUS       40924_26704        4157 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKIKOlacZKm, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4157)
  AUTHORS   Sabri S, Steen JA, Bongers M, Nielsen LK, Vickers CE.
  TITLE     Knock-in/Knock-out (KIKO) vectors for rapid integration of large DNA
            sequences, including whole metabolic pathways, onto the Escherichia 
            coli chromosome at well-characterised loci
  JOURNAL   Microb. Cell Fact. 12 (1), 60 (2013)
  PUBMED    23799955
REFERENCE   2  (bases 1 to 4157)
  AUTHORS   Steen JA, Sabri S, Bongers M, Nielsen LK, Vickers CE.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-JAN-2013) Australian Institute for Bioengineering and 
            Nanotechnology, University of Queensland, Crn College and Cooper 
            Rds, St Lucia, QLD 4072, Australia
REFERENCE   3  (bases 1 to 4157)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4157)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2013"; volume: "12"; issue: "1"; pages: "60"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-JAN-2013) Australian Institute for Bioengineering and 
            Nanotechnology, University of Queensland, Crn College and Cooper 
            Rds, St Lucia, QLD 4072, Australia"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(35..418)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(491..1348)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(1349..1453)
                     /label=AmpR promoter
     misc_feature    complement(1491..2001)
                     /label=partial lacZ fragment
                     /note="partial lacZ fragment"
     regulatory      2008..2039
                     /label=hairpin loop
                     /note="hairpin loop"
                     /regulatory_class="terminator"
     misc_feature    2061..2109
                     /label=FRT site for Flp/Frt recombination
                     /note="FRT site for Flp/Frt recombination"
     protein_bind    complement(2062..2109)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             2470..3261
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     protein_bind    complement(3455..3488)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     regulatory      3601..3640
                     /label=hairpin loop
                     /note="hairpin loop"
                     /regulatory_class="terminator"
     misc_feature    complement(3646..4150)
                     /label=partial lacZ fragment
                     /note="partial lacZ fragment"