Cas9-NAT
(Plasmid #64329)

• Purpose: Cas9 expression cassette carried by a yeast single-copy episome plasmid
• Depositing Lab: Yong-Su Jin
• Publication: Zhang et al Appl Environ Microbiol. 2014 Dec;80(24):7694-701. doi: 10.1128/AEM.02310-14. Epub 2014 Oct 3.

Backbone

• Vector backbone: pRS414-TEF1p-Cas9-CYC1t
• Backbone manufacturer: George Church lab (Addgene Plasmid #43802)
• Backbone size w/o insert (bp): 9232
• Total vector size (bp): 10446
• Vector type: Yeast Expression, CRISPR
• Selectable markers: Nourseothricin(clonNat)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: Top10 also works
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: Human-optimized S. pyogenes Cas9 with Clonnat marker gene expression cassette
• Insert Size (bp): 1206

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: SnaBI (not destroyed)
• 3′ cloning site: MfeI (not destroyed)
• 5′ sequencing primer: tctagagcggccgcTACGTAgatctgtttagcttgccttg
• 3′ sequencing primer: tctagagcggccgcCAATTGcgttttcgacactggatggc

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Plasmid p414-TEF1p-Cas9-CYC1t from George Church lab purchased from Addgene
• Articles Citing this Plasmid:
  • 15 References

Terms and Licenses

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