pHis-TEV-Nsp15-A81V
(Plasmid #219525)

• Purpose: E. coli plasmid (T7lac promoter) for full-length, SARS CoV-2 Nsp15-A81V (uridine-specific nidoviral endoribonuclease, expression-optimized gene) with N-terminal TEV protease cleavable His6-tag
• Depositing Lab: Debra Hansen
• Publication: Sonowal et al Protein Sci. 2025 Jun;34(6):e70115. doi: 10.1002/pro.70115.

Backbone

• Vector backbone: pET-11b
• Backbone manufacturer: purchased from GenScript
• Backbone size w/o insert (bp): 5644
• Total vector size (bp): 6760
• Modifications to backbone: None. GenScript cloned the expression-optimized insert into NdeI and BamHI sites of pET-11b.
• 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: Nsp15-A81V
• Alt name: NendoU-A81V
• Alt name: non-structural protein 15, A81V
• Alt name: uridylate-specific endoribonuclease, A81V
• Species: Severe acute respiratory syndrome coronavirus 2
• Insert Size (bp): 1037
• Mutation: A81V (C to T)
• Entrez Gene: N (a.k.a. GU280_gp10)
• Tag / Fusion Protein:
  • TEV protease cleavable His6-tag (N terminal on insert)

Resource Information

• Supplemental Documents:
  • pHis-TEV-Nsp15-A81V DTH GENBANK.txt
• A portion of this plasmid was derived from a plasmid made by: Purchased from GenScript the expression-optimized insert in the pET-11b backbone

Terms and Licenses

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

Depositor Comments

Protein sequence is MHHHHHHSSGVDLGTENLYFQSNAM + Nsp15-A81V (amino acid residues 6453-6798 of YP_009724389, with A81V mutation).
Confers ampicillin-resistance.
Backbone vector is pET-11b from GenScript.
Mutation design by Nirupa Nagaratnam.