pCMV-T7-AsCas12a-P2A-EGFP (RTW2861)
(Plasmid #160140)

• Purpose: CMV and T7 promoter expression plasmid for human codon optimized AsCas12a with a c-terminal NLS(nucleoplasmin), 3x HA tag, and P2A-EGFP
• Depositing Labs: Keith Joung, Benjamin Kleinstiver
• Publication: Walton et al Nat Protoc. 2021 Feb 5. pii: 10.1038/s41596-020-00465-2. doi: 10.1038/s41596-020-00465-2.

Backbone

• Vector backbone: JDS246 (Plasmid #43861)
• Vector type: Mammalian Expression ; 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 AsCas12a with NLS(nucleoplasmin)-3xHA-P2A-EGFP
• Alt name: RTW2861
• Species: Synthetic
• Promoter: CMV and T7
• Tag / Fusion Protein:
  • NLS(nucleoplasmin)-3xHA-P2A-EGFP (C terminal on insert)

Cloning Information

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

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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