pOSIP-KC vector (V012437)

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LOCUS       pOSIP-KC.        6970 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Prokaryotic one-step cloning and chromosomal integration vector 
            encoding a kanamycin resistance marker and the phage phi-C31 
            integrase.
ACCESSION   .
VERSION     .
KEYWORDS    pOSIP-KC
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6970)
  AUTHORS   St-Pierre F, Cui L, Priest DG, Endy D, Dodd IB, Shearwin KE.
  TITLE     One-step cloning and chromosomal integration of DNA.
  JOURNAL   ACS Synth Biol 2013;2:537-41.
  PUBMED    24050148
REFERENCE   2  (bases 1 to 6970)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 6970)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth 
            Biol"; date: "2013"; volume: "2"; pages: "537-41"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     Because of the toxic ccdB gene, this vector must be propagated in a 
            permissive strain such as DB3.1(TM).
FEATURES             Location/Qualifiers
     source          1..6970
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..75
                     /label=MCS 1
                     /note="MCS 1"
                     /note="multiple cloning site, part 1"
     CDS             142..444
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     rep_origin      477..1065
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1125..1145
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1200..1227
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(1276..1307)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription
                     terminator"
     protein_bind    1384..1417
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(1427..2137)
                     /label=lambda repressor (ts)
                     /note="temperature-sensitive variant of the phage lambda
                     repressor"
     CDS             2272..4086
                     /label=phage phi-C31 integrase
                     /note="integrase from phage phi-C31"
     CDS             4224..5036
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      5161..5549
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     protein_bind    5560..5607
                     /label=FLP recombinase from the Saccharomyces
                     cerevisi
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     protein_bind    complement(5676..5775)
                     /label=phage phi-C31 attP
                     /note="attachment site of phage phi-C31"
     terminator      complement(5860..6106)
                     /label=lambda tL3 terminator
                     /note="transcription terminator tL3 from phage lambda"
     terminator      6428..6514
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      6611..6697
                     /gene="Escherichia coli rrnB"
                     /label=Escherichia coli rrnB terminator
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      complement(6797..6840)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"