pET28a-HA-CPDSalI
(Plasmid #38254)

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

Backbone

• Vector backbone: pET28a
• Backbone manufacturer: Novagen
• Backbone size (bp): 6029
• Modifications to backbone: HA epitope tag and Vibrio cholerae MARTX toxin (rtxA, vc1451) cysteine protease domain (CPD) (amino acids 3440-3650) cloned into SalI and XhoI sites in backbone to create fusions with target protein of interest
• Vector type: Bacterial Expression
• Promoter: T7
• Tags / Fusion Proteins:
  • HA (C terminal on backbone)
  • Vibrio cholerae MARTX toxin cysteine protease domain (CPD) (C terminal on backbone)
  • 6xHis (C terminal on backbone)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 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 NcoI and SalI sites to create HA-tagged fusion with Vibrio cholerae MARTX toxin cysteine protease domain (CPD). HA epitope tag will remain on target protein after cleavage.