pCMV-T7-ABEmax(7.10)-SpG-P2A-EGFP (RTW4562)
(Plasmid #140002)

• Purpose: CMV and T7 promoter expression plasmid for human codon optimized ABEmax(7.10) A-to-G base editor with SpG(D10A/D1135L/S1136W/G1218K/E1219Q/R1335Q/T1337R) and P2A-EGFP
• Depositing Lab: Benjamin Kleinstiver
• Publication: Walton et al Science. 2020 Apr 17;368(6488):290-296. doi: 10.1126/science.aba8853. Epub 2020 Mar 26.

Backbone

• Vector backbone: pCMV_ABEmax_P2A_GFP
• Backbone manufacturer: David Liu (Addgene Plasmid 112101)
• Vector type: Mammalian Expression, CRISPR ; in vitro transcription; T7 promoter

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: human codon optimized ABEmax(7.10) SpCas9 variant named SpG with P2A-EGFP
• Alt name: RTW4562
• Species: Synthetic
• Mutation: nSpCas9=D10A; SpG=D1135L/S1136W/G1218K/E1219Q/R1335Q/T1337R
• Promoter: CMV and T7
• Tag / Fusion Protein:
  • P2A-EGFP (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: oBK6928-CCAAGTCTCCACCCCATTGACG
• 3′ sequencing primer: oBK219-GGGAGTGGCACCTTCCAGGGTC

Resource Information

• Supplemental Documents:
  • RTW4562
• Articles Citing this Plasmid:
  • 6 References

Terms and Licenses

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

Depositor Comments

The depositing laboratory has released the equivalent ABE8e versions of this enzyme, which they have found to show much higher editing efficiency. See https://www.addgene.org/browse/article/28228617/ for the plasmids.