pNIC-Bio3
(Plasmid #250992)

• Purpose: (Empty Backbone) Bacterial expression of proteins with N-terminal His6-TEV tag and C-terminal Avitag for in vivo (or in vitro) biotinylation with BirA. Kanamycin resistant, T7-Lac driven expression
• Depositing Lab: Opher Gileadi
• Publication: Keates et al N Biotechnol. 2012 Jun 15;29(5):515-25. doi: 10.1016/j.nbt.2011.10.007. Epub 2011 Oct 19.

Backbone

• Vector backbone: pET28
• Modifications to backbone: Added N-terminal His6-TEV tag, a negative selection insert including SacB, which confers sensitivity to 5% sucrose; and a C-terminal Avi tag.
• Vector type: Bacterial Expression
• Promoter: T7-Lac
• Tags / Fusion Proteins:
  • His6 (N terminal on backbone)
  • TEV cleavage site (N terminal on backbone)
  • Avi tag (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: For expression, grow in BL21(DE3) strains or T7Express.
• Copy number: High Copy

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: T7F
• 3′ sequencing primer: T7R

Terms and Licenses

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

Depositor Comments

Cloning into BsaI sites by Ligation-independent cloning (LIC) or restriction cloning.