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Permits
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These permits may be required for shipping this product:
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Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
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Vector Information
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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
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Applications
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encodes an epitope tag for protein isolation or monitoring expression vector vector permitting construction of fusion proteins
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Comments
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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.
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References
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Hakes DJ, Dixon JE. New vectors for high level expression of recombinant proteins in bacteria. Anal. Biochem. 202: 293-298, 1992. PubMed: 1519755
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Shipping Information
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Freeze dried plasmid in E. coli
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