pAC84-pCR8-dCas9
(Plasmid #48218)

• Purpose: dCas9 on Gateway donor vector pCR8/GW/TOPO. Note: This is not for expression. It has to be transferred to a gateway destination vector for expression
• Depositing Lab: Rudolf Jaenisch
• Publication: Cheng et al Cell Res. 2013 Aug 27. doi: 10.1038/cr.2013.122.

Backbone

• Vector backbone: pCR8/GW/TOPO
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 2799
• Total vector size (bp): 7023
• Vector type: CRISPR ; Gateway Donor

Growth in Bacteria

• Bacterial Resistance(s): Spectinomycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: dCas9(D10A;H840A)
• Alt name: dCas9
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 4224
• Mutation: D10A;H840A
• Tag / Fusion Protein:
  • HA Tag (N terminal on insert)

Cloning Information

• Cloning method: TOPO Cloning
• 5′ sequencing primer: M13F
• 3′ sequencing primer: M13R

Resource Information

• Supplemental Documents:
  • Genbank File
• A portion of this plasmid was derived from a plasmid made by: A two-step fusion PCR was performed to amplify Cas9 Nickase ORF from the pX335 vector (Addgene: 42335) and incorporate H840A mutation on the internal primers
• Articles Citing this Plasmid:
  • 2 References

Terms and Licenses

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

Depositor Comments

For more information including protocols and
updates, please go to http://www.crispr-on.org