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M50 Super 8x TOPFlash
(Plasmid #12456)

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Available to Academic and Nonprofits Only

Backbone

  • Vector backbone
    pTA-Luc
  • Backbone manufacturer
    Clontech
  • Backbone size w/o insert (bp) 4871
  • Vector type
    Luciferase

Growth in Bacteria

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

Sequence Information

Gene/Insert

  • Gene/Insert name
    TCF/LEF binding sites
  • Alt name
    beta catenin reporter
  • Insert Size (bp)
    120
  • Entrez Gene
    Ctnnb1 (a.k.a. Bfc, Catnb, Mesc)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site MluI (unknown if destroyed)
  • 3′ cloning site MluI (unknown if destroyed)
  • 5′ sequencing primer RVPrimer3
  • 3′ sequencing primer LucNRev
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    The idea for the construct is from Hans Clevers lab, who designed the original TOPflash.
  • Terms and Licenses

Depositor Comments

This is a luciferase reporter of beta-catenin-mediated transcriptional activation. In HEK cells, maximal activation of this reporter is ~100-fold (activation by Wnt) up to ~1,000-fold (activation by phosphorylation mutants of beta-catenin). The appropriate control plasmid is clone M51, Super8XFOPflash, which has mutant TCF/LEF binding sites.

This construct was made by Ajamete Kaykas in the Moon lab. The backbone is the pTA-Luc vector of Clontech, which provides a minimal TA viral promoter driving expression of the firefly luciferase gene (see company publications for details). 7 TCF/LEF binding sites were cloned into the Mlu1 site of this vector (7 copies of: AGATCAAAGGgggta, with TCF/LEF binding site in CAP letters, and a spacer in lower case, separating each copy of the TCF/LEF site).

Note: This plasmid was published as M50 Super 8x TOPFlash, but the plasmid actually contains 7 TCF/LEF sites.

How to cite this plasmid

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:

    M50 Super 8x TOPFlash was a gift from Randall Moon (Addgene plasmid # 12456)
  • For your References section:

    Zebrafish prickle, a modulator of noncanonical Wnt/Fz signaling, regulates gastrulation movements. Veeman MT, Slusarski DC, Kaykas A, Louie SH, Moon RT. Curr Biol. 2003 Apr 15. 13(8):680-5. 10.1016/S0960-9822(03)00240-9 PubMed 12699626