pCMV_T7-PE2-Nuclease
(Plasmid #171997)

• Purpose: Sequence provided for PE-Nuclease mRNA transcription, suitable for microinjection
• Depositing Lab: Paul Thomas
• Publication: Adikusuma et al Nucleic Acids Res. 2021 Sep 17. pii: 6371974. doi: 10.1093/nar/gkab792.

Backbone

• Vector backbone: pCMV-PE2
• Vector type: Mammalian Expression
• Selectable markers: Puromycin

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: CMV_T7-Cas9-RT
• Species: Synthetic; Cas9 is from S. pyogenes; M-MLV RT is from the Moloney murine leukemia virus
• Mutation: GC to CA point mutation in SpCas9 to restore nuclease function
• Promoter: CMV for Cas9, U6 for gRNAs

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pcmv_t7-pe2-nuclease.gb

Terms and Licenses

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

Depositor Comments

Full sequence https://benchling.com/s/seq-3W561bliRmUNCaGRVBPO.