pGTM_SUMO_PLpro_C111S
(Plasmid #233740)

• Purpose: Expresses SARS-CoV-2 Nsp3 residues 746-1063 for the purification of PLpro protease with a C111S active site mutation that abolishes protease activity. Following Sumo Protease cleavage no nonnative res
• Depositing Lab: Gaetano Montelione
• Publication: De Falco et al STAR Protoc. 2025 Sep 19;6(3):103952. doi: 10.1016/j.xpro.2025.103952. Epub 2025 Jul 18.

Backbone

• Vector backbone: pET15Sumo2_NESG
• Backbone manufacturer: Novagen/Northeast Structural Genomics Consortium
• Backbone size w/o insert (bp): 5737
• Total vector size (bp): 6691
• Modifications to backbone: n N-terminal 6X-His tag fused to the Smt3 gene (Sumo) of Saccharomyces cerevisiae as a purification/solubility enhancement cassette. BsaI sites are included in the cloning polylinker for scarless cloning of the SARS-CoV2 3CLpro and the production of a native, mature protein after cleavage from the Sumo fusion by Sumo protease (Ulp1).
• 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: PLpro
• Alt name: Nsp3
• Species: SARS-CoV-2
• Insert Size (bp): 957
• Mutation: Cys111Ser active site mutation of the Papain-Like Protease
• GenBank ID: WQM42602.1 PRO_0000449637
• Entrez Gene: ORF1ab (a.k.a. GU280_gp01)
• Promoter: T7 lac
• Tag / Fusion Protein:
  • 6xHis-Sumo (N terminal on backbone)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: T7 Forward
• 3′ sequencing primer: T7 Reverse

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Genscript was contracted to produce this construct

Terms and Licenses

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