pAsCpf1(TATV)(BB) (pY221)
(Plasmid #89354)

• Purpose: Expresses humanized AsCpf1 TATV PAM variant and crRNA guide
• Depositing Lab: Feng Zhang
• Publication: Gao et al Nat Biotechnol. 2017 Jun 5. doi: 10.1038/nbt.3900.

Backbone

• Vector backbone: pUC ori vector
• Backbone size w/o insert (bp): 4328
• Total vector size (bp): 8435
• Vector type: Mammalian Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): NEB Stable
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: Cpf1 RVR variant
• Alt name: AsCas12a, RVR variant
• Species: Synthetic; Acidaminococcus sp.
• Insert Size (bp): 4107
• Mutation: S542R/K548V/N552R
• Promoter: CBh
• Tags / Fusion Proteins:
  • NLS (N terminal on insert)
  • NLS-3xHA (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: AGGGATGGTTGGTTGGTGGG

Resource Information

• Supplemental Documents:
  • U6(BbsI)-CBh-NLS-AsCpf1(RVR)-NLS-3xHA.gb
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

This variant also recognizes TTTV and RATR PAMs (R = A or G).

Contains a U6-crRNA expression cassette (BbsI restriction sites for guide cloning).

For more information on Zhang lab CRISPR plasmids, please refer to www.addgene.org/crispr/zhang/.