pBV-Luc wt MBS1-4
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||16564||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerVogelstein Lab
- Backbone size w/o insert (bp) 4867
Vector typeMammalian Expression, Luciferase
Growth in Bacteria
Copy numberHigh Copy
Gene/Insert namec-MYC binding sites
SpeciesH. sapiens (human)
Entrez GeneMYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
/ Fusion Protein
- Luciferase (C terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site KpnI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer RVprimer3
- 3′ sequencing primer LucNrev (Common Sequencing Primers)
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 GCACGTGACC GCCACCCATG CGCTGAGGGG CGGACAGGAG GTGCTTCGAC TGGGAGGAGG GCGAAGAGTG TAAGGGGGCG GAGGGGCGAT GGCAGCC-3' [Oligo B, for wild-type 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.
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:pBV-Luc wt MBS1-4 was a gift from Bert Vogelstein (Addgene plasmid # 16564 ; http://n2t.net/addgene:16564 ; RRID:Addgene_16564)
For your References section:Identification of CDK4 as a target of c-MYC. Hermeking H, Rago C, Schuhmacher M, Li Q, Barrett JF, Obaya AJ, O'Connell BC, Mateyak MK, Tam W, Kohlhuber F, Dang CV, Sedivy JM, Eick D, Vogelstein B, Kinzler KW. Proc Natl Acad Sci U S A. 2000 Feb 29. 97(5):2229-34. 10.1073/pnas.050586197 PubMed 10688915