URL: http://www.addgene.org/79526
Proper Citation: RRID:Addgene_79526
Bacterial Resistance: Kanamycin
Vector Backbone Description: Backbone Marker:SGC; Backbone Size:7649; Vector Backbone:pET28-MKH8SUMO; Vector Types:Bacterial Expression; Bacterial Resistance:Kanamycin
Comments: Please see http://www.thesgc.org/sites/default/files/toronto_vectors/pET28-MKH8SUMO.pdf for more information. The pET28-MKH8SUMO vector was derived from expression plasmid pET28a-LIC (SGC). It is used for T7 promoter driven expression of recombinant proteins with the addition of an N-terminal fusion tag containing 8X His followed by a thrombin cleavage site, a SUMO, and a TEV cleavage site. Two lysines were encoded after the Met start site. Two stop codons are included in the vector at the downstream cloning site. Insertion of DNA sequence into the cloning/expression region is performed using ClonTech In-Fusion enzyme mediated directional recombination between complementary 15 nucleotide DNA sequences at the ends of the insert (PCR product) and BseRI/BsaI linearized vector. Insertion of target sequence involves replacement of a SacB gene stuffer sequence, which provides for negative selection of the original plasmid on 5% sucrose.The 15-bp primer extensions are compatible with those for pET28-MHL, such that the PCR inserts prepared for pET28-MHL can be directly used for cloning into pET28-MKH8SUMO.
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Source: Addgene