pUC-H1-gRNA
(Plasmid #61089)

• Purpose: CRISPR/Cas9 system gRNA cloning
• Depositing Lab: Shenglin Huang
• Publication: Zheng et al Biotechniques. 2014 Sep 1;57(3):115-24. doi: 10.2144/000114196. eCollection 2014.

Backbone

• Vector backbone: pUC19
• Backbone size w/o insert (bp): 2636
• Vector type: Mammalian Expression, CRISPR ; /Cas9 gRNA cloning vector

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: H1 promoter; gRNA sequences
• Species: H. sapiens (human)
• Insert Size (bp): 346
• Promoter: H1

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: M13-F20
• 3′ sequencing primer: M13R

Resource Information

• Supplemental Documents:
  • CRISPR gRNA Cloning Protocol
• Addgene Notes:
  • Addgene diagnostic digest
• A portion of this plasmid was derived from a plasmid made by: NS
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

As Ampicillin gene and H1 promoter contain BsaI sites, we mutated the sequence of Amp gene (G1601C, not change amino acid), and mutated H1 promoter from GAGACC to GAGGACC to eliminate the BsaI internal site.