pJ609-dCas9-KR-puro
(Plasmid
#158478)
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PurposeExpress nuclease dead Cas9 fused to KillerRed chromophore in mammalian cells
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Depositing Lab
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Depositing OrganizationDana-Farber Cancer Institute
Located in the United States of America -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 158478 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 158478-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 158478-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepJ609
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Backbone manufacturerDNA2.0
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Vector typeMammalian Expression, CRISPR
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Selectable markersPuromycin
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 namedCas9-KR
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SpeciesSynthetic
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Insert Size (bp)4899
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MutationCas9m4 (D10A+D839A+H840A+N863A)
- Promoter CMV
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Tag
/ Fusion Protein
- KillerRed (C terminal on insert)
Cloning Information
- Cloning method Unknown
- 5′ sequencing primer Unknown
- (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byDNA2.0
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
Cas9m4 sequence taken from Addgene plasmid #47316; KillerRed sequence from Evrogen (http://evrogen.com/products/KillerRed/KillerRed_Detailed_description.shtml) KillerRed is fused to Cas9m4 with a 5 amino acid linker.
DNA (Catalog # 158478-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
-
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.
DNA (Catalog # 158478-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:
pJ609-dCas9-KR-puro was a gift from Brendan Price (Addgene plasmid # 158478 ; http://n2t.net/addgene:158478 ; RRID:Addgene_158478) -
For your References section:
Site-specific targeting of a light activated dCas9-KillerRed fusion protein generates transient, localized regions of oxidative DNA damage. House NCM, Parasuram R, Layer JV, Price BD. PLoS One. 2020 Dec 17;15(12):e0237759. doi: 10.1371/journal.pone.0237759. eCollection 2020. 10.1371/journal.pone.0237759 PubMed 33332350
Map uploaded by the depositor.
