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PA-DAD-dark
(Plasmid #53096)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 53096 Plasmid sent as bacteria in agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pTriEx4
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 5238
  • Vector type
    Mammalian Expression, Bacterial Expression, Insect Expression

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    LOV(dark)-6aa-DAD
  • Alt name
    mVenus - 3x GS linker - LOV2-Jα (404-546) - 3x GS linker - DAD (1177-1222)
  • Alt name
    LOV2 "dark state” (closed conformation) mutant
  • Alt name
    pTriEx-mVenus-LOV(dark)-6aa-DAD
  • Species
    M. musculus (mouse); Avena sativa
  • Mutation
    L47F and C49S in Venus; C450M mutation in LOV2
  • Tags / Fusion Proteins
    • 6x His (N terminal on backbone)
    • mVenus (N terminal on backbone)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site HindIII (not destroyed)
  • 5′ sequencing primer pTriExUP
  • 3′ sequencing primer pTriExDOWN
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Expresses photoactivatable (PA) Diaphanous auto-regulatory domain (DAD) consisting of a fusion of mVenus, dark LOV2 Jα protein domain of Avena sativa Phototropin1, and Diaphanous auto-regulatory domain of mouse Diaphanous-related formin-1 (mDia1)

Please note that the C450M mutation in LOV2 creates a "dark state" mutant. See Yi et al., ACS Synth Biol. 2014 Nov 21;3(11):788-95 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:

    PA-DAD-dark was a gift from Klaus Hahn & Ronen Zaidel-Bar (Addgene plasmid # 53096)
  • For your References section:

    An optogenetic tool for the activation of endogenous diaphanous-related formins induces thickening of stress fibers without an increase in contractility. Rao MV, Chu PH, Hahn KM, Zaidel-Bar R. Cytoskeleton (Hoboken). 2013 Jul;70(7):394-407. doi: 10.1002/cm.21115. Epub 2013 May 24. 10.1002/cm.21115 PubMed 23677607