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pTriEx-RhoA FLARE.sc Biosensor T19N
(Plasmid #12152)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 12152 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pTriEx
  • Backbone manufacturer
    Novagen
  • Backbone size w/o insert (bp) 5000
  • 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
    RhoA Biosensor T19N
  • Alt name
    6xHis-RBD-CFP-Linker-Citrine-RhoA T19N
  • Alt name
    RhoA
  • Species
    H. sapiens (human)
  • Insert Size (bp)
    2400
  • Mutation
    RhoA T19N (dominant-negative mutation)*
  • Entrez Gene
    RHOA (a.k.a. ARH12, ARHA, RHO12, RHOH12)
  • Tag / Fusion Protein
    • 6xHis (N terminal on insert)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NcoI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer TriExUP
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Protocols for production and use of Hahn lab biosensors and photactivatable proteins can be found on their web page, at the URL below. If you have suggestions for the protocols or have any questions, please feel free to contact the Hahn lab. Good luck with your experiments!

Hahn lab biosensor protocol: http://www.hahnlab.com/tools/index.html

Please reference plasmid 12150 pTRIEX-RhoA Biosensor WT for complete sequence of biosensor

Please note: ECFP sequence contains K27R and N165H amino acid changes

*There is also a previously unreported A88T mutation in this plasmid. It is not known to affect function.

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:

    pTriEx-RhoA FLARE.sc Biosensor T19N was a gift from Klaus Hahn (Addgene plasmid # 12152 ; http://n2t.net/addgene:12152 ; RRID:Addgene_12152)
  • For your References section:

    Spatiotemporal dynamics of RhoA activity in migrating cells. Pertz O, Hodgson L, Klemke RL, Hahn KM. Nature. 2006 Apr 20. 440(7087):1069-72. 10.1038/nature04665 PubMed 16547516