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pSBbi-RP-CoV2/RBD-puro
(Plasmid #161793)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 161793 Standard format: Plasmid sent in bacteria as agar stab 1 $85
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Backbone

  • Vector backbone
    pSBbi-RP (Addgene Plasmid #60513)
  • Backbone manufacturer
    Eric Kowarz
  • Backbone size w/o insert (bp) 6625
  • Total vector size (bp) 7136
  • 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 RBD
  • Species
    SARS-CoV-2
  • Insert Size (bp)
    735
  • 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′
    • (Common Sequencing Primers)

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

The SARS-CoV-2 RBD insert was designed by Jorge L. Arias-Arias from Universidad de Costa Rica ([email protected])

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 RBD protein. This protein can be recovered from the cell culture medium by His-tag purification.

The recombinant RBD is suitable for COVID-19 serology testing and monitoring of humoral responses in vaccinated population.

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:

    pSBbi-RP-CoV2/RBD-puro was a gift from Jorge Luis Arias-Arias (Addgene plasmid # 161793 ; http://n2t.net/addgene:161793 ; RRID:Addgene_161793)

  • For your References section:

    Stable production of recombinant SARS-CoV-2 receptor-binding domain in mammalian cells with co-expression of a fluorescent reporter and its validation as antigenic target for COVID-19 serology testing. Arias-Arias JL, Molina-Castro SE, Monturiol-Gross L, Lomonte B, Corrales-Aguilar E. Biotechnol Rep (Amst). 2023 Mar;37:e00780. doi: 10.1016/j.btre.2022.e00780. Epub 2022 Dec 31. 10.1016/j.btre.2022.e00780 PubMed 36619904
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