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pMitoTimer
(Plasmid #52659)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 52659 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pDsRed2-Mito
  • Backbone manufacturer
    Clontech
  • Backbone size w/o insert (bp) 3959
  • Total vector size (bp) 4658
  • Modifications to backbone
    We constructed pMitoTimer by inserting the BamHI-NotI fragments of pTimer-1 (Clontech) into pDsRed2-Mito (Clontech) vector digested with BamHI and NotI using T4 ligase (New England Biolabs) following DNA isolation by using the Qiaex II gel extraction kit (Qiagen). The mitochondrial targeting sequence of the human cytochrome c oxidase subunit VIII (Cox8) gene from pDsRed2-Mito was then fused to the mutant DsRed1-E5 on the N-terminus.
  • Vector type
    DsRed
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    DsRed1-E5
  • Alt name
    Timer
  • Species
    Discosoma
  • Insert Size (bp)
    681
  • Promoter CMV
  • Tag / Fusion Protein
    • human cytochrome c oxidase subunit VIII mitochondrial targeting sequence (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site NotI (not destroyed)
  • 5′ sequencing primer CMV-F
  • 3′ sequencing primer SV40pA-R (5'-GAAATTTGTGATGCTATTGC-3')
  • (Common Sequencing Primers)

Resource 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:

    pMitoTimer was a gift from Zhen Yan (Addgene plasmid # 52659 ; http://n2t.net/addgene:52659 ; RRID:Addgene_52659)
  • For your References section:

    A novel MitoTimer reporter gene for mitochondrial content, structure, stress and damage in vivo. Laker RC, Xu P, Ryall KA, Sujkowski A, Kenwood BM, Chain KH, Zhang M, Royal MA, Hoehn KL, Dirscoll M, Adler PN, Wessells RJ, Saucerman JJ, Yan Z. J Biol Chem. 2014 Mar 18. 10.1074/jbc.M113.530527 PubMed 24644293