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Addgene

pFLAG-137.144S Tetherin
(Plasmid #41072)

Full plasmid sequence is not available for this item.

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 41072 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pFLAG-Hsp90α
  • Backbone manufacturer
    Len Neckers Laboratory (NCI, Bethesda, MD) (Koga et al. 2006, PMID: 16844778)
  • Backbone size w/o insert (bp) 5400
  • Total vector size (bp) 6000
  • Modifications to backbone
    From pcDNA3: in-frame N-terminal FLAG epitope added, Hsp90α cds added. From pFLAG-Hsp90α: Hsp90α cds replaced with Tetherin cds by BamHI and XhoI digestion.
  • 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
    Tetherin
  • Alt name
    BST-2
  • Alt name
    bone marrow stromal cell antigen 2
  • Alt name
    CD317
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    550
  • Mutation
    137S, 144S
  • GenBank ID
    NM_004335.2 NP_004326.1
  • Entrez Gene
    BST2 (a.k.a. CD317, HM1.24, TETHERIN)
  • Promoter CMV
  • Tag / Fusion Protein
    • FLAG (N terminal on backbone)

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 BGH-rev
  • (Common Sequencing Primers)

Resource Information

  • Addgene Notes
  • A portion of this plasmid was derived from a plasmid made by
    Plasmid pFLAG-Hsp90α from the laboratory of Len Neckers (NCI, Bethesda, MD) (Koga et al. 2006, PMID: 16844778)

Terms and Licenses

  • Academic/Nonprofit Terms
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

This plasmid was generated by cloning of tetherin from plasmid pCMV-HA.Tetherin (Addgene plasmid #41068) into the BamHI and XhoI sites of pFLAG-Hsp90α to generate pFLAG-Tetherin (Addgene plasmid #41070). Serine substitutions at positions aa137 and aa144 were then generated by Quik Change II Site-directed Mutagenesis Kit (Strategene).

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:

    pFLAG-137.144S Tetherin was a gift from Paul Spearman (Addgene plasmid # 41072 ; http://n2t.net/addgene:41072 ; RRID:Addgene_41072)
  • For your References section:

    The tetherin/BST-2 coiled-coil ectodomain mediates plasma membrane microdomain localization and restriction of particle release. Hammonds J, Ding L, Chu H, Geller K, Robbins A, Wang JJ, Yi H, Spearman P. J Virol. 2012 Feb;86(4):2259-72. Epub 2011 Nov 30. 10.1128/JVI.05906-11 PubMed 22130541