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CRISPR/Cas Plasmids (S.pyogenes)

Gateway cloning plasmid set Streptococcus pyogenes
 
  • pDe-CAS9 (Nuclease) - sequence information
    • Cas9 expression vector (Cas9 driven by the constitutive Ubi4-2 promotor from parsley)
    • Binary Gateway destination vector (contains ccdB gene!)
    • Bacterial selection: Spectinomycin (100 mg/l)
    • Plant selection: DL-Phosphinothricin (PPT, 6 mg/l)
      • BAR resistance cassette can easily be exchanged by HindIII
      • Please ask for pDe-CAS9-GentR if you are interested in gentamycin selection instead of PPT.
         
  • pDe-CAS9-D10A (Nickase) - sequence information
    • Cas9 nickase expression vector based on the pDe-CAS9 backbone
    • Plant selection: Kanamycin (30 mg/l)
       
  • pEn-Chimera (sgRNA) - sequence information
    • sgRNA coding vector (sgRNA can easily be customized for any target site of interest; sgRNA is driven by the Arabidopsis U6-26 promoter)
    • Gateway entry vector
    • Bacterial selection: Ampicillin (100 mg/l)
       
  • pEn-C1.1 (sgRNAs for paired nickases or paired nucleases) - sequence information
    • sgRNA coding vector based on pEn-Chimera BUT allows cloning of up to 3 different sgRNAs into pDe-CAS9
    • Gateway entry vector
    • Bacterial selection: Ampicillin (100 mg/l)

       
Conventional cloning plasmid set Streptococcus pyogenes
 
  • pCAS9-TPC (Nuclease) - sequence information
    • Cas9 expression vector (Cas9 driven by the constitutive Ubi4-2 promotor from parsley)
    • Bacterial selection: Spectinomycin (100 mg/l)
    • Plant selection: DL-Phosphinothricin (PPT, 6 mg/l)
       
  • pChimera (sgRNA) - sequence information
    • sgRNA coding vector (sgRNA can easily be customized for any target site of interest; sgRNA is driven by the Arabidopsis U6-26 promoter)
    • sgRNA is not subcloned into pCAS9-TPC via Gateway cloning but via AvrII