|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||8827||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerNew England Biolabs
- Backbone size w/o insert (bp) 6700
Vector typeBacterial Expression
Growth in Bacteria
Bacterial Resistance(s)Chloramphenicol and Ampicillin
Growth instructionsShifting the temperature from 37C to 30C during induction maximizes the yield of soluble TEV protease. Ampicillin for p793; Chloramphenicol for pRIL.
Copy numberLow Copy
Gene/Insert nameTEV protease, S219V mutant
Alt nametobacco etch virus protease
Insert Size (bp)750
MutationS219V mutation (improves stability)
/ Fusion Proteins
- His (N terminal on insert)
- polyarginine (C terminal on insert)
- MBP (with TEV) (N terminal on insert)
- 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)
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.
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