NLS-nCas9-NLS-P2A-EGFP (pRZ70)
(Plasmid #123616)

• Purpose: CMV promoter expression plasmid for codon-optimized bpNLS-32AA linker-hSpCas9n(D10A)-bpNLS-P2A-EGFP (ABEmax control without TadA domains).
• Depositing Lab: Keith Joung
• Publication: Grunewald et al Nature. 2019 Apr 17. pii: 10.1038/s41586-019-1161-z. doi: 10.1038/s41586-019-1161-z.

Backbone

• Vector backbone: CMV
• Backbone manufacturer: pCMV-BE1 (Addgene #73019)
• Backbone size w/o insert (bp): 3399
• Total vector size (bp): 8535
• Vector type: Mammalian 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: NLS-nCas9-NLS-P2A-EGFP
• Alt name: bpNLS-32AA linker-hSpCas9n(D10A)-bpNLS-P2A-EGFP
• Species: Synthetic; Streptococcus pyogenes
• Insert Size (bp): 5136
• Mutation: D10A in SpCas9
• GenBank ID:
• Promoter: CMV

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CGCAAATGGGCGGTAGGCGTG
• 3′ sequencing primer: TAGAAGGCACAGTCGAGG

Resource Information

• Supplemental Documents:
  • pRZ70.gb
• A portion of this plasmid was derived from a plasmid made by: Liu Lab (ABEmax-P2A-EGFP, Addgene #112101)
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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