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

• Depositing Lab: Bert Vogelstein
• Publication: Hermeking et al Proc Natl Acad Sci U S A. 2000 Feb 29. 97(5):2229-34.

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

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
  • Luciferase Limited Use Label License
• Industry Terms:
  • Not Available to Industry

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.