pCMV-PE7-P2A-hMLH1dn
(Plasmid #222997)

• Purpose: Mammalian expression of SpCas9 PE7 prime editor with P2A-human MLH1dn (codon optimized)
• Depositing Lab: Brittany Adamson
• Publication: Yan et al Nature. 2024 Apr 3. doi: 10.1038/s41586-024-07259-6.

Backbone

• Vector backbone: pCMV
• Backbone size w/o insert (bp): 3402
• Total vector size (bp): 12795
• Vector type: Mammalian Expression

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: PE7-P2A-hMLH1dn
• Alt name: PEmax-La(SSB)1-194-P2A-hMLH1dn
• Species: H. sapiens (human), Synthetic; SpCas9 is from Streptococcus pyogenes; MMLV_RT is from the Moloney murine leukemia virus; La(SSB)1-194 is from Homo sapiens
• Insert Size (bp): 9393
• Mutation: Deletion of residues 754-756 in MLH1 as described in https://doi.org/10.1016/j.cell.2021.09.018
• Promoter: CMV
• Tags / Fusion Proteins:
  • SV40 bpNLS (N terminal on insert)
  • SV40 bpNLS (C terminal on insert)
  • c-Myc NLS (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV Forward, T7
• 3′ sequencing primer: BGHR

Resource Information

• Supplemental Documents:
  • pCMV-PE7-P2A-hMLH1dn.gb

Terms and Licenses

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