Botanisches Institut

CRISPR/Cas12a Plasmids (L. bacterium ND2006)

Gateway cloning plasmid set Lachnospiraceae bacterium ND2006

pDeLb-Cas12a (Nuclease) sequence information

  • Cas12a expression vector (Cas12a driven by the constitutive Ubi4-2 promoter from parsley)
  • Binary Gateway destination vector (contains ccdB gene!)
  • Bacterial selection: Spectinomycin (100 mg/l)
  • Plant selection: DL-Phosphinothricin (PPT, 6 mg/l)

 

pDeEC-Lb-Cas12a (Nuclease) sequence information

  • Cas12a expression vector (Cas12a driven by the egg-cell specific promoter (fusion of EC1.1 promoter and EC1.2 enhancer) with rbcSE9 terminator)
  • Binary Gateway destination vector (contains ccdB gene!)
  • Bacterial selection: Spectinomycin (100 mg/l)
  • Plant selection: Gentamycin (60 mg/l)

 

pDettLb-Cas12a (Nuclease) sequence information

  • Cas12a expression vector (LbCas12a-D156R driven by the constitutive Ubi4-2 promoter from parsley)
  • Binary Gateway destination vector (contains ccdB gene!)
  • Bacterial selection: Spectinomycin (100 mg/l)
  • Plant selection: Gentamycin (60 mg/l)

 

pEnRZ-Lb-Chimera (crRNA) sequence information

  • crRNA coding vector (crRNA can easily be customized for any target site of interest; crRNA is driven by the Arabidopsis U6-26 promoter)
  • Gateway entry vector
  • Bacterial selection: Ampicillin (100 mg/l)