pcDNA3/Myc-DNMT3A
(Plasmid #35521)

• Purpose: Mammalian expression of human DNMT3A with myc tag
• Depositing Lab: Arthur Riggs
• Publication: Chen et al J Cell Biochem. 2005 Aug 1;95(5):902-17.

Backbone

• Vector backbone: pcDNA3
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5400
• Modifications to backbone: pcDNA3/Myc was constructed by Zhaoxia Chen (see Comments section for details). DNMT1 was cloned into the pcDNA3/Myc vector.
• 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: DNMT3A
• Species: H. sapiens (human)
• Insert Size (bp): 2739
• GenBank ID: AF067972
• Entrez Gene: DNMT3A (a.k.a. DNMT3A2, HESJAS, M.HsaIIIA, TBRS)
• Promoter: CMV
• Tag / Fusion Protein:
  • Myc (N terminal on backbone)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: BamHI (not destroyed)
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: BGH-rev

Resource Information

• Supplemental Documents:
  • MCS doc
  • Annotated GenBank file
• Articles Citing this Plasmid:
  • 32 References

Terms and Licenses

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

Depositor Comments

The multiple cloning region of the pcDNA3 vector has been modified. In the modified vector, referred to as pcDNA3/Myc, the region of the multiple cloning site located between the HindIII and NotI sites was replaced by a DNA segment containing a Kozak consensus sequence, an ATG start site and the Myc epitope sequence followed by EcoRI and BamHI restriction sites (see supplemental document). The construction of pcDNA3/Myc has been described in Chen, Z.X., et al. J Cell Biochem, 95: 902-917.

The full-length cDNA for human DNMT3A was cloned into EcoRI and BamHI sites of pcDNA3/Myc. Constructed by Zhaoxia Chen