pHPK1212
(Plasmid #8834)

• Depositing Lab: David Waugh
• Publication: Nallamsetty et al Protein Expr Purif. 2004 Nov . 38(1):108-15.

Backbone

• Vector backbone: pMal-C2
• Backbone manufacturer: New England Biolabs
• Backbone size w/o insert (bp): 6700
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): BL21(DE3)
• Growth instructions: Shifting the temperature from 37C to 30C during induction maximizes the yield of soluble MBP-TVMV protease fusion protein.
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: tobacco vein mottling virus protease, catalytic domain
• Alt name: TVMV protease
• Species: tobacco vein mottling virus
• Insert Size (bp): 700
• Tag / Fusion Protein:
  • MBP (N terminal on insert)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ cloning site: N/A (Gateway) (not destroyed)
• 3′ cloning site: N/A (Gateway) (not destroyed)
• 5′ sequencing primer: N/A
• 3′ sequencing primer: M13-F20

Terms and Licenses

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

Depositor Comments

pHPK1212 overproduces the catalytic domain of tobbaco vein mottling virus (TVMV) protease fused to the C-terminus of E. coli maltose-binding protein. The specificity of TVMV protease is distinct from that of TEV protease. The canonical recognition site for TVMV protease is ETVRFQS, with cleavage occurring between Q and S. Shifting the temperature from 37C to 30C during induction maximizes the yield of soluble MBP-TVMV protease fusion protein. The TVMV protease open reading frame on pHPK1212 was constructed from synthetic oligonucleotides and has been codon-optimized for expression in E. coli.