pSSA-RPG
(Plasmid
#85932)
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PurposeConstitutively express DsRed and conditionally express Puro and EGFP. Used to test transfection efficiency and enrich genetically modified cells
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Depositing Lab
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Depositing OrganizationNorthwest Agriculture and Forestry University
Located in China -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 85932 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 85932-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 85932-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepXL-Bac II
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Vector typeMammalian Expression ; dual-reporter surrogate
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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 namepPB-CMV-DsRed-polyA-CAG-SSA. Puro-T2A-EGFP
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Alt nameDsRed, CAG-SSA. Puro-T2A-EGFP
Cloning Information
- Cloning method Unknown
- 5′ sequencing primer pCAG-F (5'-GCAACGTGCTGGTTATTGTG-3')
- 3′ sequencing primer SV40pA-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.
Depositor Comments
Puromycin and EGFP will not be expressed until the interrupted puro gene is repaired by single strand annealing (SSA) pathway after a nuclease generates double strand breaks (DSBs).
Nuclease target sequence can be annealed and inserted into the backbone between BamHI and NotI sites using restriction enzymes or Gibson cloning methods.
DNA (Catalog # 85932-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
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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 # 85932-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:
pSSA-RPG was a gift from Zhiying Zhang (Addgene plasmid # 85932 ; http://n2t.net/addgene:85932 ; RRID:Addgene_85932) -
For your References section:
Dual-reporter surrogate systems for efficient enrichment of genetically modified cells. Ren C, Xu K, Liu Z, Shen J, Han F, Chen Z, Zhang Z. Cell Mol Life Sci. 2015 Jul;72(14):2763-72. doi: 10.1007/s00018-015-1874-6. Epub 2015 Mar 1. 10.1007/s00018-015-1874-6 PubMed 25725802
Map uploaded by the depositor.
