pCOLADuet-1_nsp10 (SARS-CoV-2)
(Plasmid #169158)

• Purpose: To express SARS-CoV-2 nsp10 in E. coli
• Depositing Lab: John Diffley
• Publication: Canal et al Biochem J. 2021 Jul 16;478(13):2445-2464. doi: 10.1042/BCJ20210198.

Backbone

• Vector backbone: pCOLADuet-1
• Total vector size (bp): 4107
• Vector type: Bacterial Expression

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: nsp10
• Alt name: untagged nsp10
• Alt name: SARS-CoV-2 nsp10 cofactor
• Species: SARS-CoV-2
• Mutation: Codon optimised for E. coli
• GenBank ID:
• Entrez Gene: ORF1ab (a.k.a. GU280_gp01)
• Promoter: T7
• Tag / Fusion Protein:
  • Methionine (N terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: ATGCGACTCCTGCATTAG
• 3′ sequencing primer: ATGCGGCCGTGTACAATAC

Resource Information

• Supplemental Documents:
  • pCOLADuet-1_nsp10 (SARS-CoV-2)_169158.gb

Terms and Licenses

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

Depositor Comments

Unlike SARS-CoV-2 nsp10 that gets expressed in the context of a larger polypeptide, this SARS-CoV-2 nsp10 gets expressed from a 5' ATG, and thus might contain a methionine at the N-terminus that is not present in the nsp5-cleaved nsp10 in cells.