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Addgene

pBV-Luc mut MBS1-3
(Plasmid #16568)

Full plasmid sequence is not available for this item.

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 16568 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pBV-Luc
  • Backbone manufacturer
    Vogelstein Lab
  • Backbone size w/o insert (bp) 4867
  • 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
    c-MYC binding sites
  • Species
    H. sapiens (human)
  • Mutation
    Mutant MBS1-3
  • Entrez Gene
    MYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
  • Tag / Fusion Protein
    • Luciferase (C terminal on backbone)

Cloning Information

  • 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)

Terms and Licenses

Trademarks:
  • Zeocin® is an InvivoGen trademark.

Depositor Comments

To generate reporter constructs, the following oligonucleotides were used:
5'-CCGGTACCGG GTTGTGGCAG CCAGTCACCT GCCCGCCGCG TAGCCACACC TCTGCTCCTC AGAGCAATGT CAAGCGGTCA CCTGTGATAG CAACAGATCA CCTGGCTGCC ATCGCCCCTC-3' [Oligo C , for mutant MBS1-3];
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.

How to cite this plasmid ( Back to top)

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 mut MBS1-3 was a gift from Bert Vogelstein (Addgene plasmid # 16568 ; http://n2t.net/addgene:16568 ; RRID:Addgene_16568)
  • 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