dCas9-NS3-NLS/VPR
(Plasmid #112244)

• Purpose: Encodes the drug-preservable Cas9-NS3-NLS/VPR, which can be used to activate gene expression when combined with an NS3 inhibitor and appropriately designed sgRNA.
• Depositing Lab: John Ngo
• Publication: Tague et al Nat Methods. 2018 Jul;15(7):519-522. doi: 10.1038/s41592-018-0042-y. Epub 2018 Jul 2.

Backbone

• Vector backbone: pcDNA3
• Backbone size w/o insert (bp): 5500
• Total vector size (bp): 12204
• Modifications to backbone: Hygromycin resistance
• Vector type: Mammalian Expression
• Selectable markers: Hygromycin

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: dCas9-NS3-NLS/VPR
• Species: S. pyogenes
• Insert Size (bp): 6700
• Mutation: NS3 mutant T54A
• Promoter: CMV
• Tags / Fusion Proteins:
  • HA (C term on NS3)
  • AU1 (N term on NS3)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV-Forward
• 3′ sequencing primer: BGH-Reverse

Resource Information

• Supplemental Documents:
  • pcdna3_dcas9-ns3-vpr-ape.gb
• A portion of this plasmid was derived from a plasmid made by: dCas9-VPR was adapted from Chavez et al., 2015 (Addgene plasmid #63798) and NS3 was adapted from the Roger Tsien lab.

Terms and Licenses

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