pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro
(Plasmid #141391)

• Purpose: Lentiviral expression of SARS-CoV-2 nucleocapsid (N) protein; transient expression and generate lentivirus
• Depositing Lab: Nevan Krogan
• Publication: Gordon et al BioRxiv March 2020

Backbone

• Vector backbone: pLVX-EF1alpha-IRES-Puro
  (Search Vector Database)
• Backbone manufacturer: Takara Bio
• Backbone size w/o insert (bp): 8825
• Vector type: Mammalian Expression, Lentiviral
• Selectable markers: Puromycin

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin
• Growth Temperature: 30°C
• Growth Strain(s): NEB Stable
• Copy number: Unknown

Gene/Insert

• Gene/Insert name: SARS-CoV-2-N
• Species: SARS-CoV-2
• Mutation: human codon optimized
• Entrez Gene: N (a.k.a. GU280_gp10)
• Promoter: EF1alpha
• Tag / Fusion Protein:
  • 2xStrep (C terminal on insert)

Cloning Information

• Cloning method: Unknown

Resource Information

• Supplemental Documents:
  • pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro.gb
• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Industry MTA

Information for Lentiviral Prep (Catalog # 141391-LV)

Purpose

Ready-to-use Lentiviral Prep particles produced from pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro (#141391). In addition to the viral particles, you will also receive purified pLVX-EF1alpha-SARS-CoV-2-N-2xStrep-IRES-Puro plasmid DNA.

Lentiviral particles carrying SARS-CoV-2 nucleocapsid (N) protein. This virus is used to make stable cell lines expressing the SARS-CoV-2 nucleocapsid protein.

Delivery

• Volume: 1mL
• Titer: ≥ 1x10⁶ TU/mL
• Pricing: $100 USD for preparation of 1mL virus + $30 USD for plasmid.
• Storage: Store at -80℃. Thaw just before use and keep on ice.
• Shipment: Viral particles are shipped frozen on dry ice. Plasmid DNA (≥ 200ng) will also be included in the shipment.

Viral Production & Use

• Packaging Plasmids: psPAX2 (plasmid #12260)
• Envelope: pMD2.G (plasmid #12259)
• Buffer: OptiPro
• Selectable Marker: Puromycin

Biosafety

Requestor is responsible for compliance with their institution's biosafety regulations. Lentivirus is generally considered BSL-2. AAV is generally considered BSL-1, but may require BSL-2 handling depending on the insert. Biosafety Guide

Terms and Licenses

• Academic/Nonprofit Terms:
  • Terms of Use for Viral Vectors
• Industry Terms:
  • Not Available to Industry

Viral Quality Control

Titering Method:

• ddPCR assay: 293T cells were transduced with serial dilutions of 141391-LV, harvested several days later, and genomic DNA was isolated. Copies of RRE were measured and normalized to RPP30.

Notes:

• PCR confirmation of insert: PCR was carried out on the viral preparation with primers targeting the pLenti backbone and the insert. The PCR product was visualized on an agarose gel for size confirmation.

Visit our viral production page for more information.