pT7-PEmax for IVT
(Plasmid #178113)

• Purpose: Template for in vitro transcription of PEmax
• 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: pT7
• Modifications to backbone: Inactivated T7 promoter to prevent transcription from plasmid
• Vector type: Template for in vitro transcription

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
• 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): 6393
• Mutation: Detailed in manuscript
• Promoter: T7 (inactivated)
• 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: Ligation Independent Cloning
• 5′ sequencing primer: CMV-forward
• 3′ sequencing primer: BGH-reverse

Resource Information

• Supplemental Documents:
  • PEmax for IVT.gb
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

Addgene's NGS results identified a one base pair mismatch in the RT region of the plasmid relative to the depositor's reference sequence. The mismatch does not impact the amino acid sequence and is not predicted to impact plasmid function.