pETGEXCT (ATCC® 87059)

Applications: encodes removable tag for protein isolationexpression vectorvector permitting RNA synthesis in vitrovector permitting construction of fusion proteins  /  Depositors: AD Sharrocks

Permits and Restrictions

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Designations pETGEXCT
Depositors AD Sharrocks
Biosafety Level 1
Vector Information
Size (kb): 5.3000001907348630
Vector: pETGEXCT (plasmid)
Promoters: Promoter T7 (phi10)
Construction: pET3d, pGEX2T
Marker(s):ampR
Construct size (kb): 5.300000190734863
Features: marker(s): ampR
other: thrombin cleavage site I
other: thrombin cleavage site II
promoter: T7 (phi10)
replicon: pMB1
restriction site: BamHI
restriction site: NcoI
restriction site: SacI
coding sequence: GST
Applications
encodes removable tag for protein isolation
expression vector
vector permitting RNA synthesis in vitro
vector permitting construction of fusion proteins
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--5.6; NcoI--5.6; SstI--5.6.
The thrombin cleavage sites produced in the protein enable the release of the fused protein from the GST tag.
The order of the major features in the plasmid is: T7 promoter - NcoI - SacI - thrombin I encoding site - GST coding sequence - thrombin II encoding site - BamHI - ampR - pMB1 ori.
The plasmid yield is rather low and thrombin cleavage of the fusion proteins can be inefficient.
Expression vector allowing production of an N-terminal or C-terminal fusion to glutathione S-transferase (GST) for affinity purification of the recombinant protein.
Media ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Sharrocks AD. A T7 expression vector for producing N- and C-terminal fusion proteins with glutathione S-transferase. Gene 138: 105-108, 1994. PubMed: 8125285

Andrew D Sharrocks, personal communication

Shipped frozen
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