pET28-MKH8SUMO
(Plasmid #79526)

• Purpose: (Empty Backbone) Bacterial expression plasmid used for T7 promoter driven expression of recombinant proteins with N-terminal 8X His, thrombin cleavage site, SUMO, and TEV cleavage site.
• Depositing Lab: Cheryl Arrowsmith
• Publication: SGC Toronto Q3 Q4 2015, Q1 2016 (unpublished)

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, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low 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

• Articles Citing this Plasmid:
  • 6 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

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.