pGEX-KN (ATCC® 77332)

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-KN
Depositors DJ Hakes, JE Dixon
Biosafety Level 1
Vector Information
Size (kb): 4.9720001220703130
Vector: pGEX-KN (plasmid)
Promoters: Promoter tac
Construction: pGEX-1
Marker(s):ampR
Construct size (kb): 4.972000122070313
Features: marker(s): ampR
other: thrombin cleavage site
promoter: tac
replicon: pMB1
repressor gene: lacIq
MCS: NotI 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): NotI--5.0; EcoRI/PstI--4.0, 1.0; BamHI/EcoRV--3.2, 1.8.
Cloning into this vector requires amplification of the gene using oligonucleotides prepared as in the reference and encoding the first 4 amino acids of a thrombin recognition sequence.
Expression vector for rapid purification of fusion proteins that contain no amino terminal extensions after thrombin cleavage. The amino acid after the initiator methionine must be charged.
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 a NotI site.
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 plasmid in E. coli

Product Sheet
Product Sheet
Product Sheet
Attachments Vector map for 77332