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PurposeCatalytic domain [SET] of mouse Ezh2 fused to N-terminus of dCas9; pCDNA3 vector backbone, mammalian expression
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Depositing Lab
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Depositing OrganizationUniversity of California, Davis
Located in United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 100087 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 100087-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 100087-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA3.3-TOPO
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Backbone manufacturerThermoFisher
- Backbone size w/o insert (bp) 5407
- Total vector size (bp) 10723
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Vector typeMammalian Expression, CRISPR
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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 nameEzh2[SET]
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Alt nameKMT6
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SpeciesM. musculus (mouse)
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Insert Size (bp)795
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MutationEzh2[SET] includes aa 482-746
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GenBank IDNP_031997.2
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Entrez GeneEzh2 (a.k.a. Enx-1, Enx1h, KMT6, mKIAA4065)
- Promoter CMV
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Tag
/ Fusion Protein
- 3XFLag-NLS-Ezh2[SET]-dCas9-NLS
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer CMV-F
- 3′ sequencing primer polyA TK-R
- (Common Sequencing Primers)
Resource Information
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.
DNA (Catalog # 100087-D50)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 50 μg
- Concentration 1 μg/μL
- Pricing $495 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
DNA (Catalog # 100087-D100)
Purpose
Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.
Delivery
- Amount 100 μg
- Concentration 1 μg/μL
- Pricing $585 USD
- Storage 4 ℃ (short-term) or -20 ℃ (long-term)
Terms and Licenses
-
Academic/Nonprofit Terms
-
Industry Terms
- Not Available to Industry
Quality Control
Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.
Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.
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:
Ezh2[SET]-dCas9 was a gift from David Segal (Addgene plasmid # 100087 ; http://n2t.net/addgene:100087 ; RRID:Addgene_100087) -
For your References section:
dCas9-based epigenome editing suggests acquisition of histone methylation is not sufficient for target gene repression. O'Geen H, Ren C, Nicolet CM, Perez AA, Halmai J, Le VM, Mackay JP, Farnham PJ, Segal DJ. Nucleic Acids Res. 2017 Sep 29;45(17):9901-9916. doi: 10.1093/nar/gkx578. 10.1093/nar/gkx578 PubMed 28973434
Map uploaded by the depositor.
