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PurposeExpresses RNase H1-EGFP- dCas9 fusion protein in human cells
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Depositing Lab
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Depositing OrganizationUniversity of Toronto
Located in Canada -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 139835 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 139835-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 139835-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA4/TO
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Backbone manufacturerLife Technologies/Thermo Fisher
- Backbone size w/o insert (bp) 5078
- Total vector size (bp) 10922
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Vector typeMammalian Expression
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Selectable markersZeocin
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)NEB Stable
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameRNase H1-EGFP-dCas9
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Alt nameRNase H1-EGFP-deadCas9
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SpeciesRNase H1 (H. sapiens), dCas9 (S. pyogenes)
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Insert Size (bp)5844
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MutationdCas9 protein contains two mutations: D10A and H840A
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GenBank IDRNase H1 (BT019670.1) Cas9 (AAK33936.1)
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Entrez GeneRNASEH1 (a.k.a. H1RNA, PEOB2, RNH1)
- Promoter tet-inducible CMV
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NotI (not destroyed)
- 3′ cloning site XbaI (not destroyed)
- 5′ sequencing primer CMV forward
- 3′ sequencing primer BGH reverse
- (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 # 139835-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 # 139835-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:
RED was a gift from Karim Mekhail (Addgene plasmid # 139835 ; http://n2t.net/addgene:139835 ; RRID:Addgene_139835) -
For your References section:
Nucleolar RNA polymerase II drives ribosome biogenesis. Abraham KJ, Khosraviani N, Chan JNY, Gorthi A, Samman A, Zhao DY, Wang M, Bokros M, Vidya E, Ostrowski LA, Oshidari R, Pietrobon V, Patel PS, Algouneh A, Singhania R, Liu Y, Yerlici VT, De Carvalho DD, Ohh M, Dickson BC, Hakem R, Greenblatt JF, Lee S, Bishop AJR, Mekhail K. Nature. 2020 Sep;585(7824):298-302. doi: 10.1038/s41586-020-2497-0. Epub 2020 Jul 15. 10.1038/s41586-020-2497-0 PubMed 32669707
Map uploaded by the depositor.
Map uploaded by the depositor.
