pET22b-CPDBamHI-Leu
(Plasmid #38256)

• Purpose: (Empty Backbone)
• Depositing Labs: Matthew Bogyo, Aimee Shen
• Publication: Shen et al PLoS One. 2009 Dec 2;4(12):e8119.

Backbone

• Vector backbone: pET22b
• Backbone manufacturer: Novagen
• Backbone size (bp): 6114
• Modifications to backbone: Vibrio cholerae MARTX toxin (rtxA, vc1451) cysteine protease domain (CPD) (amino acids 3444-3650) cloned into BamHI and XhoI sites in backbone to create fusions with target protein of interest
• Vector type: Bacterial Expression
• Promoter: T7
• Tags / Fusion Proteins:
  • Vibrio cholerae MARTX toxin cysteine protease domain (CPD) (C terminal on backbone)
  • 6xHis (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: T7 promoter
• 3′ sequencing primer: T7 terminator

Terms and Licenses

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

Depositor Comments

For protein purification, target protein of interest should be cloned into NdeI and BamHI sites to create fusion with Vibrio cholerae MARTX toxin cysteine protease domain (CPD)

Note that the P1 Leu for cleavage must be encoded in the 3' cloning primer of the target gene.