pBS-Hsp70-Cas9
(Plasmid #46294)

• Purpose: A codon-optimized Cas9 nuclease under the control of the Drosophila hsp70 promoter.
• Depositing Labs: Melissa Harrison, Kate O'Connor-Giles, Jill Wildonger
• Publication: Gratz et al Genetics. 2014 Apr;196(4):961-71. doi: 10.1534/genetics.113.160713. Epub 2014 Jan 29.

Backbone

• Vector backbone: pBluescript-KS(+)
• Backbone manufacturer: Stratagene
• Backbone size w/o insert (bp): 2958
• Vector type: Insect 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: codon optimized Cas9
• Alt name: SpCas9
• Mutation: Silent change from C to A at bp 834 of insert
• Promoter: Hsp70
• Tag / Fusion Protein:
  • 3x Flag (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NotI (unknown if destroyed)
• 3′ cloning site: NotI (unknown if destroyed)
• 5′ sequencing primer: T7
• 3′ sequencing primer: T3

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Cas9 insert from Feng Zhang, Broad Institute, MIT
• Articles Citing this Plasmid:
  • 28 References

Terms and Licenses

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

Depositor Comments

This plasmid was created by subcloning the Hsp70-Cas9 containing NotI fragment from Addgene plasmid 45945 in NotI digested pBluescript-KS(+).

For more information on FlyCRISPR Plasmids please refer to: http://www.addgene.org/crispr/OConnor-Giles/

Plasmid 51019: pDsRed-attP (www.addgene.org/51019) can be for generating dsDNA donors for homology-directed repair to replace genes or other genomic sequence with an attP docking site.