BPK3079
(Plasmid #78741)

• Purpose: Human expression plasmid for AsCpf1 guide RNA (need to clone in spacer into BsmBI sites)
• Depositing Lab: Keith Joung
• Publication: Kleinstiver et al Nat Biotechnol. 2016 Jun 27. doi: 10.1038/nbt.3620.

Backbone

• Vector backbone: MLM3636
• Backbone manufacturer: Joung lab (Addgene Plasmid # 43860)
• Vector type: Mammalian Expression, CRISPR

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: AsCpf1 crRNA backbone, without spacer sequence
• Species: Synthetic
• Promoter: U6

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BsmBI (not destroyed)
• 3′ cloning site: BsmBI (not destroyed)
• 5′ sequencing primer: OS280 (5'-CAGGGTTATTGTCTCATGAGCGG-3')

Resource Information

• Supplemental Documents:
  • Annotated GenBank file
  • Joung Lab gRNA Cloning Protocol
• Articles Citing this Plasmid:
  • 4 References

Terms and Licenses

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

Depositor Comments

Use BsmBI sites to insert crRNA spacer sequence into the vector. This plasmid will express gRNA in mammalian cells for Cpf1 from Acidaminococcus sp. (strain BV3L6). Recommended to use with SQT1659 (Addgene plasmid# 78743), which expresses Acidaminococcus sp. Cpf1.

Please visit https://www.biorxiv.org/content/early/2016/06/09/057802 for bioRxiv preprint.