Addgene - pBV-Luc mut MBS4 Plasmid Data

Plasmid 16570: pBV-Luc mut MBS4

Gene/insert name:
c-MYC binding sites
Species:
H. sapiens (human)
Entrez Gene:
MYC (MRTL, bHLHe39, c-Myc)
Fusion protein or tag:
Luciferase
Terminal:
C terminal on backbone
Mutation:
Mutant MBS4
Vector backbone:
pBV-Luc
(Search Vector Database)
Backbone manufacturer:
Vogelstein Lab
Vector type:
Mammalian Expression, Luciferase
Backbone size w/o insert (bp):
4867
Cloning site 5':
KpnI
Site destroyed during cloning:
No
Cloning site 3':
EcoRI
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
Sequence: View sequences (1)
Map:
View map
Principal Investigator:
Bert Vogelstein
Terms and Licenses:
MTA
Luciferase Limited Use Label License

Comments:

To generate reporter constructs, the following oligonucleotides were used:
5'-CCGGTACCGG GTTGTGGCAG CCAGTCACGT GCCCGCCGCG TAGCCACACC TCTGCTCCTC AGAGCAATGT CAAGCGGTCA CGTGTGATAG CAACAGATCA CGTGGCTGCC ATCGCCCCTC-3' [Oligo A, for wild-type c-MYC binding sites (MBS)1-3]; 5'-ATGAATTCCG GACGTTCTGG GCAGGTGACC GCCACCCATG CGCTGAGGGG CGGACAGGAG GTGCTTCGAC TGGGAGGAGG GCGAAGAGTG TAAGGGGGCG GAGGGGCGAT GGCAGCCAGG-3' [Oligo D, for mutant MBS4].

Different combinations of oligonucleotide pairs (A+B, A+D, C+B, C+D) were annealed and converted to double-stranded fragments through one PCR cycle. These promoter fragments were subcloned into pBV-luc. Further polymerase-derived mutants were identified while sequencing the reporter constructs.

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

Article: Identification of CDK4 as a target of c-MYC. Hermeking et al (Proc Natl Acad Sci U S A. 2000 Feb 29. 97(5):2229-34. 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 16570" in your Materials and Methods section.