Addgene - M51 Super 8x FOPFlash (TOPFlash mutant) Plasmid Data

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

Gene/insert name:
mutant TCF/LEF binding sites
Alt name:
beta catenin reporter
Insert size:
134
Entrez Gene:
Ctnnb1 (Bfc, Catnb, Mesc)
Mutation:
mutant TCF/LEF binding sites
Vector backbone:
pGL3
(Search Vector Database)
Backbone manufacturer:
Promega
Vector type:
Mammalian Expression, Luciferase
Backbone size w/o insert (bp):
4839
Cloning site 5':
Asp718
Site destroyed during cloning:
No
Cloning site 3':
XmaI
Site destroyed during cloning:
No
5' sequencing primer:
RVPrimer3 List of Sequencing Primers
3' sequencing primer:
LucNRev
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Person or lab that originally
cloned the gene/insert:
The idea for the construct is from Hans Clevers lab, who designed the original TOPflash.
Sequence: View sequences (3)
Principal Investigator:
Randall Moon
Terms and Licenses:
MTA
Luciferase Limited Use Label License

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.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Zebrafish prickle, a modulator of noncanonical Wnt/Fz signaling, regulates gastrulation movements. Veeman et al (Curr Biol. 2003 Apr 15. 13(8):680-5. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 12457" in your Materials and Methods section.