pJCC_163 SpCas9 K233A/K234A/K253A/K263A
(Plasmid #179527)

• Purpose: For bacterial expression of SpCas9 K233A/K234A/K253A/K263A (helix-rolling basic patch mutant) with an N-terminal His-MBP tag
• Depositing Lab: Jennifer Doudna
• Publication: Cofsky et al Nat Struct Mol Biol. 2022 Apr;29(4):395-402. doi: 10.1038/s41594-022-00756-0. Epub 2022 Apr 14.

Backbone

• Vector backbone: p2CT
• Backbone manufacturer: UC Berkeley MacroLab
• Backbone size w/o insert (bp): 5969
• Total vector size (bp): 10073
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: SpCas9 K233A/K234A/K253A/K263A
• Species: Streptococcus pyogenes
• Insert Size (bp): 4104
• Mutation: changed lysines 233, 234, 253, and 263 to alanine
• Tags / Fusion Proteins:
  • 10X His (N terminal on backbone)
  • maltose-binding protein (N terminal on backbone)
  • TEV protease cleavage site (N terminal on backbone)

Resource Information

• Supplemental Documents:
  • pJCC_163.gb
• A portion of this plasmid was derived from a plasmid made by: Addgene plasmid 101199 (pSHS207 - Bacterial expression plasmid for WT SpCas9)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry