pETDuet-1 Hepatitis C Virus NS3/NS4A
(Plasmid #61696)

• Purpose: inducible dual expression of NS3 protease along with its cofactor NS4A
• Depositing Lab: Michael Johnson
• Publication: Lee et al PLoS One. 2013 Oct 9;8(10):e75144. doi: 10.1371/journal.pone.0075144. eCollection 2013.

Backbone

• Vector backbone: pETDuet-1
• Backbone manufacturer: EMD
• Backbone size w/o insert (bp): 5420
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: NS4A
• Species: Hepatitis C Virus
• Promoter: T7

Cloning Information for Gene/Insert 1

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: HindIII (not destroyed)
• 5′ sequencing primer: pBRrevBam

Gene/Insert 2

• Gene/Insert name: NS3
• Species: Hepatitis C Virus
• Promoter: T7
• Tag / Fusion Protein:
  • his (N terminal on insert)

Cloning Information for Gene/Insert 2

• Cloning method: Restriction Enzyme
• 5′ cloning site: NdeI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: none
• 3′ sequencing primer: T7 terminal

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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

Depositor Comments

For protein expression, grow in Rosetta cells.
25 degree induction for 14-15 hours with 0.5 mM IPTG