pKIKOarsBCm vector (V005242)

Basic Vector Information

Vector Name:
pKIKOarsBCm
Antibiotic Resistance:
Ampicillin
Length:
3771 bp
Type:
Cloning vector
Replication origin:
R6K γ ori
Source/Author:
Sabri S, Steen JA, Bongers M, Nielsen LK, Vickers CE.

pKIKOarsBCm vector Map

pKIKOarsBCm3771 bp60012001800240030003600R6K gamma oriAmpRAmpR promoterpartial arsB fragmenthairpin loopFRTCmRcat promoterFRT (minimal)hairpin looppartial arsB fragment

pKIKOarsBCm vector Sequence

LOCUS       40924_26689        3771 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKIKOarsBCm, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3771)
  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 3771)
  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 3771)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3771)
  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..3771
                     /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..2058)
                     /label=partial arsB fragment
                     /note="partial arsB fragment"
     regulatory      2063..2094
                     /label=hairpin loop
                     /note="hairpin loop"
                     /regulatory_class="terminator"
     protein_bind    complement(2117..2164)
                     /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)."
     misc_feature    2117..2158
                     /label=FRT site for Flp/Frt recombination
                     /note="FRT site for Flp/Frt recombination"
     CDS             complement(2184..2840)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(2841..2943)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     protein_bind    complement(3049..3082)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     regulatory      complement(3195..3234)
                     /label=hairpin loop
                     /note="hairpin loop"
                     /regulatory_class="terminator"
     misc_feature    complement(3240..3764)
                     /label=partial arsB fragment
                     /note="partial arsB fragment"

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