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pGTM_COV2_NSP5_004_SUMO
(Plasmid #190062)

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Item Catalog # Description Quantity Price (USD)
Plasmid 190062 Standard format: Plasmid sent in bacteria as agar stab 1 $85 *
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Backbone

  • Vector backbone
    pET15Sumo2_NESG
  • Backbone manufacturer
    Novagen/Northeast Structural Genomics Consortium
  • Backbone size w/o insert (bp) 5737
  • Total vector size (bp) 6604
  • Modifications to backbone
    An 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
    Room Temperature
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    3CLpro
  • Alt name
    SARS-CoV-2 Main Protease
  • Alt name
    Nsp5
  • Species
    SARS-CoV-2
  • Insert Size (bp)
    918
  • GenBank ID
    PRO_0000449623
  • Entrez Gene
    ORF1ab (a.k.a. GU280_gp01)
  • Promoter T7 lac
  • Tag / Fusion Protein
    • 6X-His-Sumo (N terminal on backbone)

Cloning Information

  • Cloning method Ligation Independent Cloning
  • 5′ sequencing primer T7 Forward
  • 3′ sequencing primer T7 Reverse
    • (Common Sequencing Primers)

Resource Information

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

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This construct was produced by total gene synthesis by Genscript. The DNA insert is optimized for expression in Escherichia coli.

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:

    pGTM_COV2_NSP5_004_SUMO was a gift from Gaetano Montelione (Addgene plasmid # 190062 ; http://n2t.net/addgene:190062 ; RRID:Addgene_190062)

  • For your References section:

    Protocol for production and purification of SARS-CoV-2 3CL(pro). Mazzei L, Greene-Cramer R, Bafna K, Jovanovic A, De Falco A, Acton TB, Royer CA, Ciurli S, Montelione GT. STAR Protoc. 2023 May 5;4(2):102326. doi: 10.1016/j.xpro.2023.102326. 10.1016/j.xpro.2023.102326 PubMed 37235475
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