pRSFDuet-1(nsp8-nsp7)(nsp12)
(Plasmid #165451)

• Purpose: Expresses the RNA Dependent RNA polymerase complex of SARS-CoV-2 in Escherichia Coli
• Depositing Lab: Marc Delarue
• Publication: Madru et al PLoS One. 2021 Apr 29;16(4):e0250610. doi: 10.1371/journal.pone.0250610. eCollection 2021.

Backbone

• Vector backbone: pRSFDuet-1
• Backbone manufacturer: Novagen
• Backbone size w/o insert (bp): 3829
• Modifications to backbone: the multi cloning site 1 was modified to encode a N-terminal 14xHis tag (instead of 6xHis)
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert 1

• Gene/Insert name: nsp12
• Alt name: RNA-directed RNA polymerase
• Alt name: Non-structural protein 12
• Species: Severe acute respiratory syndrome coronavirus 2
• Insert Size (bp): 2799
• GenBank ID: QIZ64668.1
• Promoter: T7

Cloning Information for Gene/Insert 1

• Cloning method: Unknown
• 5′ sequencing primer: DuetUP2
• 3′ sequencing primer: T7term

Gene/Insert 2

• Gene/Insert name: nsp8
• Alt name: Non-structural protein 8
• Species: severe acute respiratory syndrome coronavirus 2
• Insert Size (bp): 603
• GenBank ID: QIZ64668.1
• Promoter: T7
• Tag / Fusion Protein:
  • 14Histidine-TEV (N terminal on backbone)

Cloning Information for Gene/Insert 2

• Cloning method: Unknown
• 5′ sequencing primer: ACYCDuetUP1
• 3′ sequencing primer: DuetDOWN1

Gene/Insert 3

• Gene/Insert name: nsp7
• Alt name: Non-structural protein 7
• Species: severe acute respiratory syndrome coronavirus 2
• Insert Size (bp): 255
• GenBank ID: QIZ64668.1
• Promoter: T7

Cloning Information for Gene/Insert 3

• Cloning method: Unknown
• 5′ sequencing primer: ACYCDuetUP1
• 3′ sequencing primer: DuetDOWN1

Resource Information

• Supplemental Documents:
  • 20AD6JAC_RSFDuetCS1nsp87nsp12.gb
• A portion of this plasmid was derived from a plasmid made by: The open reading frames (ORF) of the nsp7, nsp8 and nsp12 genes from SARS-CoV-2 virus were synthesized commercially by geneArt (Thermo Fisher) and inserted into a modified pRSFDuet-1 (Novagen). The codon usage of the three ORFs have been optimized for expression in E. coli.
• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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