pcDNA3.3_CoV2_D18
(Plasmid #170442)

• Purpose: Express SARS-CoV-2 WT spike protein with an 18aa deletion on the C-terminal tail. Used for SARS2 pseudotyped virus production.
• Depositing Lab: David Nemazee
• Publication: Rogers et al Science. 2020 Aug 21;369(6506):956-963. doi: 10.1126/science.abc7520. Epub 2020 Jun 15.

Backbone

• Vector backbone: pcDNA3.3
• Backbone size w/o insert (bp): 5500
• Total vector size (bp): 9200
• Modifications to backbone: NA
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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: SARS-CoV-2 spike D18
• Species: SARS-CoV-2, USA-WA1/2020 strain
• Insert Size (bp): 3768
• Mutation: 18aa deletion in c-terminal tail
• GenBank ID: 43740568
• Entrez Gene: S (a.k.a. GU280_gp02, spike glycoprotein)
• Promoter: CMV
• Tag / Fusion Protein:
  • NA

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (not destroyed)
• 3′ cloning site: XhoI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: TK-pA-R

Resource Information

• Supplemental Documents:
  • SARS-CoV-2 pseudotyped virus (PSV) production protocol
• Addgene Notes:
  • Addgene Diagnostic Digest
• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

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