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pBMN DHFR(DD)-YFP
(Plasmid #29325)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 29325 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pBMN
  • Backbone manufacturer
    Garry Nolan
  • Backbone size w/o insert (bp) 6943
  • Vector type
    Retroviral
  • Selectable markers
    Hc-Red-t fluorecent protein behind IRES

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    30°C
  • Growth Strain(s)
    Top10
  • Growth instructions
    TOP10 cells. Culture at 30C to prevent recombination of LTR and lost insert. Takes >20 hours to get good culture at 30C.
  • Copy number
    Low Copy

Gene/Insert

  • Gene/Insert name
    DHFR(DD)-YFP
  • Alt name
    folA dihydrofolate reductase ( destable domain) with YFP fusion
  • Species
    E. coli
  • Insert Size (bp)
    1236
  • Mutation
    Change Arg 12 to Tyr (R12Y),Gly 67 to Ser (G67S) and Tyr 100 to Ile (Y100I)--destabilizing domain
  • GenBank ID
    AAC73159
  • Tags / Fusion Proteins
    • YFP (C terminal on insert)
    • HA (C terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site SphI (not destroyed)
  • 5′ sequencing primer pBMN 5'
  • (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:

    pBMN DHFR(DD)-YFP was a gift from Thomas Wandless (Addgene plasmid # 29325)
  • For your References section:

    A general chemical method to regulate protein stability in the mammalian central nervous system. Iwamoto M, Bjoerklund T, Lundberg C, Kirik D, Wandless TJ. Chem Biol. 2010 Sep 24. 17(9):981-8. 10.1016/j.chembiol.2010.07.009 PubMed 20851347