pcDNA3.1-hAsCpf1(TYCV) (pY210)
(Plasmid #89351)

• Purpose: Expresses humanized AsCpf1 variant that recognizes TYCV PAMs
• Depositing Lab: Feng Zhang
• Publication: Gao et al Nat Biotechnol. 2017 Jun 5. doi: 10.1038/nbt.3900.

Backbone

• Vector backbone: pcDNA3.1
• Backbone size w/o insert (bp): 5402
• Total vector size (bp): 9509
• 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, RR variant
• Alt name: AsCas12a, RR variant
• Species: Synthetic; Acidaminococcus sp.
• Insert Size (bp): 4107
• Mutation: S542R/K607R
• Promoter: CMV
• Tags / Fusion Proteins:
  • NLS (N terminal on insert)
  • NLS-3xHA (C terminal on insert)

Cloning Information

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

Resource Information

• Supplemental Documents:
  • pcDNA3.1-CMV-NLS-hAsCpf1(RR)-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 CCCC and ACCC PAMs (and, with lower efficiency, VYCV).

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