pSBbi-RP-CoV2/stSpike-puro
(Plasmid #161794)

• Purpose: Transposon-based SARS-CoV-2 secretable trimeric Spike stable expression vector for protein production
• Depositing Lab: Jorge Luis Arias-Arias
• Publication: Arias-Arias et al Biotechnol Rep (Amst). 2023 Mar;37:e00780. doi: 10.1016/j.btre.2022.e00780. Epub 2022 Dec 31.

Backbone

• Vector backbone: pSBbi-RP (Addgene Plasmid #60513)
• Backbone manufacturer: Eric Kowarz
• Backbone size w/o insert (bp): 6625
• Total vector size (bp): 10208
• Vector type: Mammalian Expression ; Transposon
• Selectable markers: Puromycin

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: SARS-CoV-2 secretable trimeric Spike
• Species: SARS-CoV-2
• Insert Size (bp): 3807
• Promoter: EF1a/RPBSA
• Tags / Fusion Proteins:
  • 6xHis (C terminal on insert)
  • dTomato (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: NcoI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: 5′-GCCTCAGACAGTGGTTCAAAG-3′
• 3′ sequencing primer: 5′-AGGCACAGTCGAGGCTGAT-3′

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Takara Bio Limited Use Label License (formerly Clontech)
• Industry Terms:
  • Not Available to Industry

Depositor Comments

The SARS-CoV-2 secretable trimeric Spike insert was designed by Jorge L. Arias-Arias from Universidad de Costa Rica ().

This vector is suitable for transient transfection, but it was intended to be co-transfected with the transposase vector pCMV(CAT)T7-SB100 (Addgene Plasmid #34879) in order to generate stable mammalian cells for the production of SARS-CoV-2 secretable trimeric Spike protein. This protein can be recovered from the cell culture medium by His-tag purification.

Please note that this plasmid is intended to be used as a DNA template of the spike gene but not as a construct for protein production. The protein produced using this plasmid is not antigenically functional.