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pRK793
(Plasmid #8827)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 8827 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

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)
    Chloramphenicol and Ampicillin, 25 & 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    BL21(DE3)-RIL
  • Growth instructions
    Shifting the temperature from 37C to 30C during induction maximizes the yield of soluble TEV protease. Ampicillin for p793; Chloramphenicol for pRIL.
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    TEV protease, S219V mutant
  • Alt name
    tobacco etch virus protease
  • Insert Size (bp)
    750
  • Mutation
    S219V mutation (improves stability)
  • Tags / Fusion Proteins
    • His (N terminal on insert)
    • polyarginine (C terminal on insert)
    • MBP (with TEV) (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site SacI (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer N/A
  • 3′ sequencing primer M13-F20
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    ATCC
  • Articles Citing this Plasmid

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Bacterial strain: E.coli BL21(DE3)-RIL (Stratagene).

pRK793 overproduces the catalytic domain of tobacco etch virus (TEV) protease in the form of an MBP fusion protein that cleaves itself in vivo to yield a TEV protease catalytic domain with an N-terminal His-tag and a C-terminal polyarginine tag.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pRK793 was a gift from David Waugh (Addgene plasmid # 8827 ; http://n2t.net/addgene:8827 ; RRID:Addgene_8827)
  • For your References section:

    Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency. Kapust RB, Tozser J, Fox JD, Anderson DE, Cherry S, Copeland TD, Waugh DS. Protein Eng. 2001 Dec . 14(12):993-1000. 10.1093/protein/14.12.993 PubMed 11809930