pαH-S-GSAS-2P-N317P
(Plasmid #196378)

• Purpose: Mammalian cell expression of SARS-CoV-2 Spike protein with mutation N317P with furin side replaced by GSAS and with 2 Proline substitution at 986 and 987
• Depositing Lab: Priyamvada Acharya
• Publication: Manrique et al iScience. 2023 Jan 20;26(1):105855. doi: 10.1016/j.isci.2022.105855. Epub 2022 Dec 26.

Backbone

• Vector backbone: pαH
• Backbone size w/o insert (bp): 4746
• Total vector size (bp): 8367
• Vector type: Mammalian Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: S-GSAS-2P-N317P
• Species: Severe acute respiratory syndrome coronavirus 2
• Insert Size (bp): 3624
• Mutation: Ectodomain (AAs 1-1208); 682-685 (furin site = RRAR) replaced with GSAS; N317P
• Entrez Gene: S (a.k.a. GU280_gp02, spike glycoprotein)
• Tags / Fusion Proteins:
  • HRV 3C cleavage site (before tags) C terminal on backbone 8X His tag C and 2X Strep-Tag II C terminal on backbone (C terminal on backbone)
  • HRV3C cleavage site, 8X His tag, 2X Strep-Tag II (C terminal on backbone)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: pCAGGS-5

Resource Information

• A portion of this plasmid was derived from a plasmid made by: A portion of this plasmid was derived from a plasmid made by Jason McLellan

Terms and Licenses

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