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

Permits and Restrictions

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Designations pGEX-KT
Depositors DJ Hakes, JE Dixon
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

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 ATCC® 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