M51 Super 8x FOPFlash (TOPFlash mutant)
(Plasmid #12457)

• Purpose: Control for beta-catenin reporter. Contains mutated TCF/LEF binding sites upstream of a luciferase reporter.
• Depositing Lab: Randall Moon
• Publication: Veeman et al Curr Biol. 2003 Apr 15. 13(8):680-5.

Backbone

• Vector backbone: pGL3
• Backbone manufacturer: Promega
• Backbone size w/o insert (bp): 4839
• Vector type: Mammalian Expression, Luciferase

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: mutant TCF/LEF binding sites
• Alt name: beta catenin reporter
• Insert Size (bp): 134
• Mutation: mutant TCF/LEF binding sites
• Entrez Gene: Ctnnb1 (a.k.a. Bfc, Catnb, Mesc)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Asp718 (not destroyed)
• 3′ cloning site: XmaI (not destroyed)
• 5′ sequencing primer: RVPrimer3
• 3′ sequencing primer: LucNRev

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.
• Articles Citing this Plasmid:
  • 191 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This is a CONTROL luciferase reporter with the TCF/LEF sites of Super8XTOPflash (construct M50, Addgene Plasmid #12456) being mutated.

There are 6 mutated TCF/LEF binding sites that were cloned into the pGL3 vector (Promega), which contains a minimal promoter followed by a firefly luciferase open reading frame. This 134bp fragment was gene synthesized and cloned into the ASP718 and XMA-1 sites of the vector.

Note: This plasmid was published as M51 Super 8x FOPFlash, but the plasmid actually contains 6 TCF/LEF sites.