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)

pDettLbCas12a-i (Nuclease) sequence information

• Cas12a expression vector (LbCas12a-D156R with introns 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)