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PurposeCas9-GFP, dual gRNA expression vector for targeted integration of repair templates into safe harbor 231 locus in chromosome 4
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Depositing Labs
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Depositing OrganizationUniversity of Washington
Located in United States of America -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 115150 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 115150-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 115150-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonecustom
- Backbone size w/o insert (bp) 2739
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Vector typeMammalian Expression, CRISPR
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 nameDual SH231 gRNAs
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Alt nameCas9-T2A-GFP
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gRNA/shRNA sequencegcctcccccatagtaccat, Gatgtgctcactgagtctga
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SpeciesStreptococcus pyogenes
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Insert Size (bp)6866
- Promoter U6, CBh
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Tags
/ Fusion Proteins
- 3xFLAG-SV40NLS (N terminal on backbone)
- GFP (C terminal on insert)
- Nucleoplasmin NLS (C terminal on backbone)
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer unknown
- (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.
Depositor Comments
Please visit https://www.biorxiv.org/content/early/2018/08/20/396390 for bioRxiv preprint.
DNA (Catalog # 115150-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 # 115150-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:
pUS2-SH231 was a gift from Raymond Monnat & Michael Phelps (Addgene plasmid # 115150 ; http://n2t.net/addgene:115150 ; RRID:Addgene_115150) -
For your References section:
New Human Chromosomal Sites with "Safe Harbor" Potential for Targeted Transgene Insertion. Pellenz S, Phelps M, Tang W, Hovde BT, Sinit RB, Fu W, Li H, Chen E, Monnat RJ Jr. Hum Gene Ther. 2019 Jul;30(7):814-828. doi: 10.1089/hum.2018.169. Epub 2019 Mar 28. 10.1089/hum.2018.169 PubMed 30793977