pET-CjCas9
(Plasmid #89754)

• Purpose: Expression of CjCas9 with His tag in E.coli
• Depositing Lab: Seokjoong Kim
• Publication: Kim et al Nat Commun. 2017 Feb 21;8:14500. doi: 10.1038/ncomms14500.

Backbone

• Vector backbone: pET28b
• Backbone manufacturer: Genescript
• Backbone size w/o insert (bp): 5400
• Total vector size (bp): 8365
• Vector type: Bacterial Expression, CRISPR

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 30°C
• Growth Strain(s): DH5alpha
• Growth instructions: Transform into BL21(DE3) for protein expression and purification purposes
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: CjCas9
• Alt name: CjeCas9
• Species: Campylobacter jejuni
• Insert Size (bp): 3048
• Promoter: T7
• Tags / Fusion Proteins:
  • 6XHis (N terminal on backbone)
  • HA (N terminal on backbone)
  • SV40 NLS (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: SacI (destroyed during cloning)
• 5′ sequencing primer: taatacgactcactatagg

Terms and Licenses

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