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PurposeExpression of codon-optimized SARS-CoV2 Nsp12 (RdRp) in mammalian cells
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Depositing Lab
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Depositing OrganizationUniversity of Pittsburgh (University, School of Medicine, and Cancer Institute)
Located in the United States of America -
Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 154008 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 * | |
| DNA | 154008-D50 | 50 μg of DNA in Tris buffer | $495 * | ||
| DNA | 154008-D100 | 100 μg of DNA in Tris buffer | $585 * | ||
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Backbone
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Vector backbonep3xFLAG-CMV-14
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Backbone manufacturerSigma
- Backbone size w/o insert (bp) 6310
- Total vector size (bp) 9159
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
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 nameCodon-optimized SARS-CoV2 Nsp12 from ORF1ab
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Alt nameSARS-CoV2 RdRp
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SpeciesSynthetic
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Insert Size (bp)2892
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GenBank IDMN985325.1
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Entrez GeneORF1ab (a.k.a. GU280_gp01)
- Promoter CMV
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Tag
/ Fusion Protein
- 3xFLAG tag (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site BamHI (not destroyed)
- 5′ sequencing primer CMV forward
- 3′ sequencing primer 3xFLAG reverse
- (Common Sequencing Primers)
Terms and Licenses
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Academic/Nonprofit Terms
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Industry Terms
Trademarks:
- Zeocin® is an InvivoGen trademark.
Depositor Comments
Insert DNA was synthesized by IDT
DNA (Catalog # 154008-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
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 # 154008-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
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:
p3xFLAG SARS-CoV2 RdRp.co was a gift from Patrick Moore (Addgene plasmid # 154008 ; http://n2t.net/addgene:154008 ; RRID:Addgene_154008) -
For your References section:
Development and characterization of a new monoclonal antibody against SARS-CoV-2 NSP12 (RdRp). Meng W, Guo S, Cao S, Shuda M, Robinson-McCarthy LR, McCarthy KR, Shuda Y, Paniz Mondolfi AE, Bryce C, Grimes Z, Sordillo EM, Cordon-Cardo C, Li P, Zhang H, Perlman S, Guo H, Gao SJ, Chang Y, Moore PS. J Med Virol. 2023 Jan;95(1):e28246. doi: 10.1002/jmv.28246. Epub 2022 Nov 2. 10.1002/jmv.28246 PubMed 36271490
Map uploaded by the depositor.
