pJ609-dCas9-KR-puro
(Plasmid #158478)

• Purpose: Express nuclease dead Cas9 fused to KillerRed chromophore in mammalian cells
• Depositing Lab: Brendan Price
• Publication: House et al PLoS One. 2020 Dec 17;15(12):e0237759. doi: 10.1371/journal.pone.0237759. eCollection 2020.

Backbone

• Vector backbone: pJ609
• Backbone manufacturer: DNA2.0
• Vector type: Mammalian Expression, CRISPR
• Selectable markers: Puromycin

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-KR
• Species: Synthetic
• Insert Size (bp): 4899
• Mutation: Cas9m4 (D10A+D839A+H840A+N863A)
• Promoter: CMV
• Tag / Fusion Protein:
  • KillerRed (C terminal on insert)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: Unknown

Resource Information

• A portion of this plasmid was derived from a plasmid made by: DNA2.0

Terms and Licenses

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

Depositor Comments

Cas9m4 sequence taken from Addgene plasmid #47316; KillerRed sequence from Evrogen (http://evrogen.com/products/KillerRed/KillerRed_Detailed_description.shtml) KillerRed is fused to Cas9m4 with a 5 amino acid linker.