M50 Super 8x TOPFlash vector (V001134)

Price Information

Cat No. Plasmid Name Availability Add to cart
V001134 M50 Super 8x TOPFlash In stock (lyophilized plasmid)

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Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.

Basic Vector Information

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.

      • Vector Name:
      • M50 Super 8x TOPFlash
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 4985 bp
      • Type:
      • Signal Pathway Reporter Vectors
      • Selection Marker:
      • Luc

M50 Super 8x TOPFlash vector Vector Map

M50 Super 8x TOPFlash4985 bp6001200180024003000360042004800minPluciferaseSV40 poly(A) signaloriAmpRAmpR promoterf1 orisynthetic polyadenylation signalpause site

Plasmid Resuspension Protocol:

1. Centrifuge at 5,000×g for 5 min.

2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.

3. Close the tube and incubate for 10 minutes at room temperature.

4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.

5.Store the plasmid at -20 ℃.

M50 Super 8x TOPFlash vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                4985 bp ds-DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4985)
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4985
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        161..192
                     /note="minP"
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             254..1906
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /note="luciferase"
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    1947..2068
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(2487..3076)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3247..4107)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4108..4212)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      4239..4694
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    4825..4873
                     /note="synthetic polyadenylation signal"
     misc_feature    4887..4978
                     /note="pause site"
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"