BPK2660
(Plasmid #70709)

• Purpose: (Empty Backbone) Human expression plasmid for SaCas9 sgRNA(84nt in length) (need to clone in spacer into BsmBI sites): U6-BsmBIcassette-Sa-sgRNA(84)
• Depositing Lab: Keith Joung
• Publication: Kleinstiver et al Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404.

Backbone

• Vector backbone: pUC19
• Backbone manufacturer: Joachim Messing (Addgene plasmid # 50005)
• Vector type: Mammalian Expression, CRISPR
• Promoter: U6

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: OS280 (5'-CAGGGTTATTGTCTCATGAGCGG-3')

Resource Information

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

Terms and Licenses

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

Depositor Comments

Use BsmBI sites to insert sgRNA spacer sequence.

The length of the sgRNA (not including the spacer sequence) is 84 nucleotides, where this truncated repeat/anti-repeat sgRNA is similar to that described previously by Ran et al., Nature 2015 (PubMed ID: 25830891) and conveys higher activity than the canonical full length sgRNA