pET-His6-SUMO-MproC145A
(Plasmid #172653)

• Purpose: Bacterial expression of SARS-CoV-2 Mpro C145A inactive mutant fused to SUMO
• Depositing Lab: Stephen Harrison
• Publication: MacDonald et al ACS Infect Dis. 2021 Sep 10;7(9):2591-2595. doi: 10.1021/acsinfecdis.1c00237. Epub 2021 Aug 26.

Backbone

• Vector backbone: pET (https://www.addgene.org/29666/)
• Backbone manufacturer: Berkeley Macrolab
• Backbone size w/o insert (bp): 4786
• Total vector size (bp): 6010
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Low Copy

Gene/Insert

• Gene/Insert name: SARS-CoV-2 Mpro
• Alt name: 3CLpro
• Species: Severe acute respiratory syndrome coronavirus 2
• Insert Size (bp): 1224
• Mutation: Catalytic Cysteine 145 mutated to Alanine
• Entrez Gene: ORF1ab (a.k.a. GU280_gp01)
• Promoter: T7
• Tags / Fusion Proteins:
  • SUMO (N terminal on insert)
  • 6xHis Tag (N terminal on backbone)

Cloning Information

• Cloning method: Ligation Independent Cloning
• 5′ sequencing primer: T7 forward
• 3′ sequencing primer: T7 reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The original Mpro gene that was mutated for this construct was received from from Linlin Zhang, University of Lubeck (https://science.sciencemag.org/content/368/6489/409)

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

SUMO fusion can be removed by cleavage with Ulp1 protease