pGEX-KT (ATCC® 77331)

Applications: encodes an epitope tag for protein isolation or monitoringexpression vectorvector permitting construction of fusion proteins  /  Depositors: DJ Hakes, JE Dixon

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Designations pGEX-KT
Depositors DJ Hakes, JE Dixon
Biosafety Level 1
Vector Information
Size (kb): 4.9840002059936520
Vector: pGEX-KT (plasmid)
Promoters: Promoter tac
Construction: pGEX-1
Marker(s):ampR
Construct size (kb): 4.984000205993652
Features: marker(s): ampR
other: thrombin cleavage site
promoter: tac
replicon: pMB1
repressor gene: lacIq
MCS: BamHI SmaI EcoRI
epitope tag: GST
Applications
encodes an epitope tag for protein isolation or monitoring
expression vector
vector permitting construction of fusion proteins
Comments
Restriction digests of the clone give the following sizes (kb): BamHI--5.0; EcoRI/PstI--4.0, 1.0; EcoRI/EcoRV--3.2, 1.8.
The resulting protein contains two additional amino acids at the amino terminus (Gly-Ser).
Expression vector for rapid purification of fusion proteins and release of proteins with fewer extraneous N-terminal amino acids.
The glutathione S-transferase (GST) fusion protein can be purified by glutathione affinity chromatography, and the desired polypeptide released from the fusion product by thrombin.
Constructed from pGEX-1 by inserting an oligonucleotide at the BamHI site which encodes the glycine "kinker" and thrombin recognition site to enhance cleavage of the fusion protein.
Media Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Growth Conditions
Temperature: 37.0°C
References

Hakes DJ, Dixon JE. New vectors for high level expression of recombinant proteins in bacteria. Anal. Biochem. 202: 293-298, 1992. PubMed: 1519755

Shipped freeze-dried
Shipping Information Freeze dried E. coli containing the plasmid
Product Sheet
Product Sheet
Product Sheet
Attachments Vector map for 77331