pD40-His/V5-c-MycT58A
(Plasmid #45598)

• Depositing Lab: Rosalie Sears
• Publication: Yeh et al Nat Cell Biol. 2004 Apr;6(4):308-18. Epub 2004 Mar 14.

Backbone

• Vector backbone: pcDNA DEST40
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 7143
• Total vector size (bp): 6841
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

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 T58A
• Alt name: c-Myc
• Alt name: Myc2
• Alt name: myelocytomatosis oncogene
• Species: M. musculus (mouse)
• Insert Size (bp): 1400
• Mutation: T58A
• Entrez Gene: Myc (a.k.a. Myc2, Niard, Nird, bHLHe39)
• Promoter: CMV
• Tags / Fusion Proteins:
  • V5 (C terminal on backbone)
  • 6xHis (C terminal on backbone)

Cloning Information

• Cloning method: Gateway Cloning
• 5′ sequencing primer: CMV-F; T7
• 3′ sequencing primer: BGH-rev

Resource Information

• A portion of this plasmid was derived from a plasmid made by: Murine c-myc cDNA amplified from plasmid CMV-Myc from Stephen R. Hann (Vanderbuilt University) (PMID: 7958908) (Murine myc cDNA originally obtained from Michael Cole, Princeton, NJ)
• Articles Citing this Plasmid:
  • 6 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Alternate plasmid names: pD40-His-c-MycT58A; pcDNA-DEST40 Myc T58A

Plasmid construction information:
Point mutations were introduced into the murine c-Myc sequence in the CMV–Myc plasmid (Hann et al., Genes Dev. 1994, PMID: 7958908) using the GeneEditor kit (Promega) per manufacturer's instructions using oligonucletide
MycT58A, 5′-CTGCTTCCCGCCCCGCCCCTGTC-3′.

The His6-tagged c-MycT58A plasmid was then created by PCR amplification with a CACC 5′ extension added to the 5′ primer for cloning into the TOPO entry clone from Invitrogen (Carlsbad, CA). This sequence was then moved into the pDEST-40 mammalian expression vector using Gateway technology (Invitrogen), creating pD40-His-c-MycT58A.

For more plasmid information (map and sequence), see the wildtype version, pD40-His/V5-c-Myc (Addgene plasmid 45597) For additional usage information, see the associated article (Yeh et al. Nat Cell Biol. 2004) as well as Arnold et al. EMBO 2009 (PMID: 19131971).