pX330-BsaIx2-DD-Cas9
(Plasmid #75380)

• Purpose: Expresses DD-SpCas9 in mammalian cells and has a cloning site for an sgRNA. The FKBP12 L106P destabilization domain allows for inducible stabilization of Cas9 through the addition of Shield-1
• Depositing Lab: Michele Calos
• Publication: Geisinger et al Nucleic Acids Res. 2016 Jan 13. pii: gkv1542.

Backbone

• Vector backbone: modified pX330
• Modifications to backbone: BbsI sites in sgRNA replaced with 2 BsaI sites
• Vector type: Mammalian Expression, CRISPR

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: DD-SpCas9
• Insert Size (bp): 4599
• Promoter: CAG
• Tag / Fusion Protein:
  • FKBP12 L106P DD (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BshTI (not destroyed)
• 3′ cloning site: NotI (not destroyed)
• 5′ sequencing primer: agaggtaagggtttaagg

Resource Information

• Supplemental Documents:
  • pX330-U6-chi_BsaIx2-CBh-DD-hSpCas9.gb

Terms and Licenses

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

Depositor Comments

DD domain tested for stabilization with 500 nM Shield-1