iCas
(Plasmid #84232)

• Purpose: Expression of SpCas9 with 4 ERT2 fusion protein and empty gRNA cassette. The activity of Cas9 can be switched on and off in human cells with 4-hydroxytamoxifen (4-HT)
• Depositing Lab: Meng How Tan
• Publication: Liu et al Nat Chem Biol. 2016 Sep 12. doi: 10.1038/nchembio.2179.

Backbone

• Vector backbone: GeneArt CRISPR Nuclease vector with OFP reporter
• Backbone size w/o insert (bp): 5081
• Vector type: Mammalian Expression
• Selectable markers: OFP expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: Cas9
• Alt name: Inducible Cas9
• Species: S. pyogenes
• Insert Size (bp): 7969
• Promoter: CMV
• Tags / Fusion Proteins:
  • ERT2-ERT2 (N terminal on insert)
  • ERT2-ERT2 (C terminal on insert)
  • 2A-OFP co-expression (C terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: AgeI/SfoI (unknown if destroyed)
• 3′ cloning site: EcoRI/XbaI (unknown if destroyed)
• 5′ sequencing primer: NA

Resource Information

• Addgene Notes:
  • Addgene diagnostic digest
  • Addgene Diagnostic Digest
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

The four copies of ERT2 are identical in sequence and may recombine occasionally. Hence, it best to use competent cells like STBL3 or NEB Stable to propagate the plasmid and to also check the plasmid size on an agarose gel.