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Addgene

Venus-tBid-4E-pEGFP-C1
(Plasmid #166743)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 166743 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pEGFP-C1
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    tBid-4E
  • Alt name
    p15 fragment of BH3-interacting domain death agonist protein, or Bcl2 like protein 10 (BCL2L10)
  • Species
    M. musculus (mouse)
  • Mutation
    4 hydrophobic residues in the BH3 region mutated to glutamic acid: I27E, L31E, I34E, M38E.
  • Entrez Gene
    Bid (a.k.a. 2700049M22Rik)
  • Promoter CMV
  • Tag / Fusion Protein
    • Venus (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site Unknown (unknown if destroyed)
  • 3′ cloning site Unknown (unknown if destroyed)
  • 5′ sequencing primer TGACGCAAATGGGCGGTAGG
  • 3′ sequencing primer TCGCCCTTTGACGTTGGAGTCCAC
  • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    tBid seqence aligns with Mus musculus BH3 interacting domain death agonist (Bid), mRNA; NCBI Reference Sequence: NM_007544.4.
  • Article Citing this Plasmid

Terms and Licenses

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

Depositor Comments

Transfection of this plasmid will express "VtBid-4E" (Venus fused to the N-terminus of the BH3-only protein, truncated Bid (tBid)) in live cells. In cells, caspase 8 cleavage of Bid produces the p15 fragment, referred to as tBid. BH3 region has hydrophobic positions H1-H4 mutated to glutamic acid (E). BH3, Mutations: I27E, L31E, I34E, M38E. When compared with EYFP, Venus fluorophore contains the following mutations: F46L, F64L, M153T, V163A and S175G. This Venus protein contains an F224R mutation to make it monomeric.

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:

    Venus-tBid-4E-pEGFP-C1 was a gift from David Andrews (Addgene plasmid # 166743 ; http://n2t.net/addgene:166743 ; RRID:Addgene_166743)
  • For your References section:

    Efficacy and specificity of inhibitors of BCL-2 family protein interactions assessed by affinity measurements in live cells. Osterlund EJ, Hirmiz N, Pemberton JM, Nougarede A, Liu Q, Leber B, Fang Q, Andrews DW. Sci Adv. 2022 Apr 22;8(16):eabm7375. doi: 10.1126/sciadv.abm7375. Epub 2022 Apr 20. 10.1126/sciadv.abm7375 PubMed 35442739