pInvRecA vector (V012461)

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LOCUS       pInvRecA.        7354 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Plasmid for transiently converting any E. coli strain to a RecA+ 
            phenotype for phage P1-mediated transduction.
ACCESSION   .
VERSION     .
KEYWORDS    pInvRecA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7354)
  AUTHORS   Sektas M, Gregorowicz M, Szybalski W.
  TITLE     Transient conversion to RecA+ phenotype to permit P1 transduction in
            any Escherichia coli recA- strains.
  JOURNAL   BioTechniques 1999;27:911-4.
  PUBMED    10572635
REFERENCE   2  (bases 1 to 7354)
  AUTHORS   Lucigen
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7354)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "BioTechniques"; date: "1999"; volume: "27"; pages: "911-4"
COMMENT     SGRef: number: 2; type: "Journal Article"
COMMENT     Addition of heated chlortetracycline induces synthesis of Flp 
            recombinase, thereby inverting the recA gene so that it is 
            expressed.
            Growth at 42(o)C inactivates the temperature-sensitive mutant Rep101
            protein, causing loss of the plasmid.
FEATURES             Location/Qualifiers
     source          1..7354
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(72..695)
                     /label=TetR
                     /note="tetracycline repressor TetR"
     promoter        711..766
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             783..2048
                     /label=FLP
                     /note="site-specific recombinase"
     protein_bind    2251..2284
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     CDS             complement(2314..3372)
                     /label=RecA
                     /note="DNA repair protein from E. coli"
     RBS             complement(3379..3401)
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(3425..3458)
                     /label=FRT (minimal)
                     /note="supports FLP-mediated excision but not integration
                     (Turan and Bode, 2011)"
     terminator      3602..3645
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3743..3829
                     /gene="Escherichia coli rrnB"
                     /label=Escherichia coli rrnB terminator
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3924..4010
                     /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(4105..4191)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(4519..5466)
                     /label=Rep101(Ts)
                     /note="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
     rep_origin      complement(5514..5736)
                     /direction=LEFT
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             complement(6361..7218)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7219..7323)
                     /label=AmpR promoter