Addgene - pRK793 Plasmid Data

Gene/insert name:
TEV protease, S219V mutant
Alt name:
tobacco etch virus protease
Insert size:
750
Fusion protein or tag:
His
Terminal:
N terminal on insert
Fusion protein or tag:
polyarginine
Terminal:
C terminal on insert
Fusion protein or tag:
MBP (with TEV)
Terminal:
N terminal on insert
Mutation:
S219V mutation
Vector backbone:
pMal-C2
(Search Vector Database)
Backbone manufacturer:
New England Biolabs
Vector type:
Bacterial Expression
Backbone size w/o insert:
6700
Cloning site 5':
SacI
Site destroyed during cloning:
No
Cloning site 3':
BamHI
Site destroyed during cloning:
No
5' sequencing primer:
N/A List of Sequencing Primers
3' sequencing primer:
M13-F20
Bacterial resistance:
Ampicillin and Chloramphenicol
Growth strain, other:
BL21(DE3)-RIL
Growth temperature (℃):
37
Growth instructions:
Shifting the temperature from 37C to 30C during induction maximizes the yield of soluble TEV protease. Ampicillin for p793; Chloramphenicol for pRIL.
High or low copy:
Low Copy
Person or lab that originally
cloned the gene/insert:
ATCC
Sequence: View sequences (2)
Principal Investigator:
David Waugh
Terms and Licenses:
MTA

Comments:

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

pRK793 overproduces the catalytic domain of tobbaco 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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Tobacco etch virus protease: mechanism of autolysis and rational design of stable mutants with wild-type catalytic proficiency. Kapust et al (Protein Eng. 2001 Dec . 14(12):993-1000. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 8827" in your Materials and Methods section.