pCMV-PEmax-P2A-GFP
(Plasmid #180020)

• Purpose: Mammalian expression of SpCas9 PEmax prime editor with P2A-EGFP marker
• Depositing Lab: David Liu
• Publication: Chen et al Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 10.1016/j.cell.2021.09.018.

Backbone

• Vector backbone: pCMV
• Vector type: Mammalian Expression
• Selectable markers: EGFP

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: PEmax-P2A-EGFP
• Alt name: SV40_bpNLS-SpCas9(R221K,N394K,H840A)-SGGSx2-SV40_bpNLS-SGGSx2-MMLV_RT(D200N, T306K, W313F, T330P, L603W)-SV40_bpNLS-cMyc_NLS
• Species: Synthetic; SpCas9 is from Streptococcus pyogenes; MMLV_RT is from the Moloney murine leukemia virus
• Insert Size (bp): 7176
• Mutation: Detailed in manuscript
• Promoter: CMV
• Tags / Fusion Proteins:
  • SV40 bpNLS (N terminal on insert)
  • SV40 bpNLS (C terminal on insert)
  • c-Myc NLS (C terminal on insert)
  • P2A-EGFP (C terminal on insert)

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: T7-forward
• 3′ sequencing primer: BGH-reverse

Resource Information

• Supplemental Documents:
  • pPC1651.gb
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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