PurposeMammalian expression of human DNMT3A with myc tag
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||35521||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5400
Modifications to backbonepcDNA3/Myc was constructed by Zhaoxia Chen (see Comments section for details). DNMT1 was cloned into the pcDNA3/Myc vector.
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
SpeciesH. sapiens (human)
Insert Size (bp)2739
Entrez GeneDNMT3A (a.k.a. DNMT3A2, HESJAS, M.HsaIIIA, TBRS)
- Promoter CMV
/ Fusion Protein
- Myc (N terminal on backbone)
- 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 (Common Sequencing Primers)
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
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:pcDNA3/Myc-DNMT3A was a gift from Arthur Riggs (Addgene plasmid # 35521 ; http://n2t.net/addgene:35521 ; RRID:Addgene_35521)
For your References section:Physical and functional interactions between the human DNMT3L protein and members of the de novo methyltransferase family. Chen ZX, Mann JR, Hsieh CL, Riggs AD, Chedin F. J Cell Biochem. 2005 Aug 1;95(5):902-17. 10.1002/jcb.20447 PubMed 15861382