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Size (kb): 5.3000001907348630
Vector: pETGEXCT (plasmid)
Promoters: Promoter T7 (phi10)
Construction: pET3d, pGEX2T
Construct size (kb): 5.300000190734863
Features: marker(s): ampR
other: thrombin cleavage site I
other: thrombin cleavage site II
promoter: T7 (phi10)
restriction site: BamHI
restriction site: NcoI
restriction site: SacI
coding sequence: GST
encodes removable tag for protein isolation
vector permitting RNA synthesis in vitro
vector permitting construction of fusion proteins
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.
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