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pET28-MKH8SUMO
(Plasmid #79526)

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Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 79526 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pET28-MKH8SUMO
  • Backbone manufacturer
    SGC
  • Backbone size (bp) 7649
  • Vector type
    Bacterial Expression
  • Tags / Fusion Proteins
    • 8xHis-Thrombin-SUMO-TEV (N terminal on backbone)
    • TEV cleavage site (N terminal on backbone)

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    None
  • Tags / Fusion Proteins
    • 8xHis-Thrombin-SUMO-TEV (N terminal on backbone)
    • TEV cleavage site (N terminal on backbone)

Resource Information

  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry

Depositor 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.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pET28-MKH8SUMO was a gift from Cheryl Arrowsmith (Addgene plasmid # 79526 ; http://n2t.net/addgene:79526 ; RRID:Addgene_79526)