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pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro
(Plasmid #141383)

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Item Catalog # Description Quantity Price (USD)
Plasmid 141383 Standard format: Plasmid sent in bacteria as agar stab 1 $85 *
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Lentiviral Prep 141383-LV Virus (1mL at titer ≥ 1x10⁶ TU/mL) and Plasmid. $180
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Backbone

  • Vector backbone
    pLVX-EF1alpha-IRES-Puro
  • 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, 100 μg/mL
  • Growth Temperature
    30°C
  • Growth Strain(s)
    NEB Stable
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    SARS-CoV-2-orf3a
  • Species
    SARS-CoV-2
  • Mutation
    human codon optimized
  • Entrez Gene
    ORF3a (a.k.a. GU280_gp03)
  • Promoter EF1alpha
  • Tag / Fusion Protein
    • 2xStrep (C terminal on insert)

Cloning Information

  • Cloning method Unknown

Resource Information

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

Information for Lentiviral Prep (Catalog # 141383-LV) ( Back to top)

Purpose

Ready-to-use Lentiviral Prep particles produced from pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro (#141383). In addition to the viral particles, you will also receive purified pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro plasmid DNA.

Lentiviral particles carrying SARS-CoV-2-orf3a. This virus is used to make stable cell lines expressing the SARS-CoV-2-orf3a.

Delivery

  • Volume 1mL
  • Titer ≥ 1x10⁶ TU/mL
  • Pricing $150 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

Viral Quality Control

Titering Method:
  • ddPCR assay: 293T cells were transduced with serial dilutions of 141383-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.

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:

    pLVX-EF1alpha-SARS-CoV-2-orf3a-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141383 ; http://n2t.net/addgene:141383 ; RRID:Addgene_141383) For viral preps, please replace (Addgene plasmid # 141383) in the above sentence with: (Addgene viral prep # 141383-LV)

  • For your References section:

    A SARS-CoV-2 protein interaction map reveals targets for drug repurposing. Gordon DE, Jang GM, Bouhaddou M, Xu J, Obernier K, White KM, O'Meara MJ, Rezelj VV, Guo JZ, Swaney DL, Tummino TA, Huttenhain R, Kaake RM, Richards AL, Tutuncuoglu B, Foussard H, Batra J, Haas K, Modak M, Kim M, Haas P, Polacco BJ, Braberg H, Fabius JM, Eckhardt M, Soucheray M, Bennett MJ, Cakir M, McGregor MJ, Li Q, Meyer B, Roesch F, Vallet T, Mac Kain A, Miorin L, Moreno E, Naing ZZC, Zhou Y, Peng S, Shi Y, Zhang Z, Shen W, Kirby IT, Melnyk JE, Chorba JS, Lou K, Dai SA, Barrio-Hernandez I, Memon D, Hernandez-Armenta C, Lyu J, Mathy CJP, Perica T, Pilla KB, Ganesan SJ, Saltzberg DJ, Rakesh R, Liu X, Rosenthal SB, Calviello L, Venkataramanan S, Liboy-Lugo J, Lin Y, Huang XP, Liu Y, Wankowicz SA, Bohn M, Safari M, Ugur FS, Koh C, Savar NS, Tran QD, Shengjuler D, Fletcher SJ, O'Neal MC, Cai Y, Chang JCJ, Broadhurst DJ, Klippsten S, Sharp PP, Wenzell NA, Kuzuoglu-Ozturk D, Wang HY, Trenker R, Young JM, Cavero DA, Hiatt J, Roth TL, Rathore U, Subramanian A, Noack J, Hubert M, Stroud RM, Frankel AD, Rosenberg OS, Verba KA, Agard DA, Ott M, Emerman M, Jura N, von Zastrow M, Verdin E, Ashworth A, Schwartz O, d'Enfert C, Mukherjee S, Jacobson M, Malik HS, Fujimori DG, Ideker T, Craik CS, Floor SN, Fraser JS, Gross JD, Sali A, Roth BL, Ruggero D, Taunton J, Kortemme T, Beltrao P, Vignuzzi M, Garcia-Sastre A, Shokat KM, Shoichet BK, Krogan NJ. Nature. 2020 Jul;583(7816):459-468. doi: 10.1038/s41586-020-2286-9. Epub 2020 Apr 30. 10.1038/s41586-020-2286-9 PubMed 32353859
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