TetO-FUW-VdC9BV
(Plasmid #62195)

• Purpose: Expresses RNA-Guided, Nuclease-Inactive VP64:dCas9-BFP:VP64—VdC9BV—Fusion Protein to Enable Transactivation of Endogenous Genes
• Depositing Lab: Kam Leong
• Publication: Chakraborty et al Stem Cell Reports. 2014 Dec 9;3(6):940-7. doi: 10.1016/j.stemcr.2014.09.013. Epub 2014 Oct 23.

Backbone

• Vector backbone: TetO-FUW
• Total vector size (bp): 13824
• Vector type: Mammalian Expression, Lentiviral

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): Stbl3
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: VP64dCas9BFPVP64
• Alt name: VdC9BV
• Species: Synthetic
• Mutation: D10A H840A (catalytically inactive) Cas9 (dCas9)
• Tag / Fusion Protein:
  • Two VP64s tagged to dCas9 fused to BFP

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NheI (not destroyed)
• 3′ cloning site: EcoRI (destroyed during cloning)
• 5′ sequencing primer: AGCTCGTTTAGTGAACCGTCAGATC
• 3′ sequencing primer: CATAGCGTAAAAGGAGCAACA

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Evrogen Limited Use Label License for FPs
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The vector backbone was from Addgene plasmid 27152 (PI Marius Wernig)
The dCas9:BFP fusion part of the insert was cloned from Addgene plasmid 44247 (PI Stanley Qi)