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PurposeEncodes for Cas9-VP64, MS2-p65-HSF1, mCherry and for the gRNA 2.0
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Depositing Lab
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Depositing OrganizationUniversity of Edinburgh
Located in the United Kingdom of Great Britain and Northern Ireland -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 99866 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 99866-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 99866-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonePiggy Bac
- Backbone size w/o insert (bp) 3961
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Vector typeMammalian Expression, CRISPR, Synthetic Biology ; dCAS9-SAM activation
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Selectable markersmCherry
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 numberLow Copy
Gene/Insert
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Gene/Insert nameUniSAM-mCherry + U6-gRNA2.0
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Alt nameCas9VP64-MS2p65HSF1-mCherry + gRNA(2xMS2Loops)
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Insert Size (bp)8388
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MutationBbsI sites in CDS were ablated by consensus mutation codon optimised for H.sapiens
- Promoter EF1a
Cloning Information
- Cloning method Gibson Cloning
- 5′ sequencing primer EXT_PB_RV--GGCACATATCAATATTATGCTCTCG
- 3′ sequencing primer EXT_PB_FW--GCAAAGCGATGACGAGCTTG
- (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 # 99866-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 # 99866-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:
PB-UniSAM was a gift from Lesley Forrester (Addgene plasmid # 99866 ; http://n2t.net/addgene:99866 ; RRID:Addgene_99866) -
For your References section:
An all-in-one UniSam vector system for efficient gene activation. Fidanza A, Lopez-Yrigoyen M, Romano N, Jones R, Taylor AH, Forrester LM. Sci Rep. 2017 Jul 25;7(1):6394. doi: 10.1038/s41598-017-06468-6. 10.1038/s41598-017-06468-6 PubMed 28743878
Map uploaded by the depositor.
