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PurposeMammalian expression of human DNMT1 with myc tag
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 36939 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA3
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5400
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Modifications to backbonepcDNA3/Myc was constructed by Zhaoxia Chen (see Comments section for details). DNMT1 was cloned into the pcDNA3/Myc vector.
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameDNMT1
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SpeciesH. sapiens (human)
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Insert Size (bp)4851
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GenBank IDX63692
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Entrez GeneDNMT1 (a.k.a. ADCADN, AIM, CXXC9, DNMT, HSN1E, MCMT, m.HsaI)
- Promoter CMV
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Tag
/ Fusion Protein
- Myc (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site NotI (not destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer BGH-rev (Common Sequencing Primers)
Resource Information
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Supplemental Documents
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Articles Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Trademarks:
- Zeocin® is an InvivoGen trademark.
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 restriction site (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 DNMT1 was cloned into EcoRI and NotI 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.
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For your Materials & Methods section:
pcDNA3/Myc-DNMT1 was a gift from Arthur Riggs (Addgene plasmid # 36939 ; http://n2t.net/addgene:36939 ; RRID:Addgene_36939) -
For your References section:
The histone methyltransferase SETDB1 and the DNA methyltransferase DNMT3A interact directly and localize to promoters silenced in cancer cells. Li H, Rauch T, Chen ZX, Szabo PE, Riggs AD, Pfeifer GP. J Biol Chem. 2006 Jul 14;281(28):19489-500. Epub 2006 May 8. 10.1074/jbc.M513249200 PubMed 16682412