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Depositing Lab
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Depositing OrganizationUniversity of North Carolina at Chapel Hill
Located in the United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 22893 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 22893-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 22893-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA3
- Backbone size w/o insert (bp) 5400
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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)DH5alpha
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameRPA2
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Alt nameRPA34
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SpeciesH. sapiens (human)
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Insert Size (bp)810
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Entrez GeneRPA2 (a.k.a. REPA2, RP-A p32, RP-A p34, RPA32)
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Tag
/ Fusion Protein
- Flag (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (unknown if destroyed)
- 3′ cloning site XbaI (unknown if destroyed)
- 5′ sequencing primer T7
- (Common Sequencing Primers)
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
There is one point mutation in the insert, but the protein expressed with this construct forms complexes with the other subunits of the RPA heterotrimer, binds other known interacting proteins (XPA, Tipin), and shows an enrichment in the chromatin fraction of cells treated with replication stress-inducing agents. So by these criteria the construct appears to be WT.
DNA (Catalog # 22893-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 # 22893-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:
pcDNA3-Flag-RPA2 was a gift from Aziz Sancar (Addgene plasmid # 22893 ; http://n2t.net/addgene:22893 ; RRID:Addgene_22893) -
For your References section:
The human Tim/Tipin complex coordinates an Intra-S checkpoint response to UV that slows replication fork displacement. Unsal-Kacmaz K, Chastain PD, Qu PP, Minoo P, Cordeiro-Stone M, Sancar A, Kaufmann WK. Mol Cell Biol. 2007 Apr . 27(8):3131-42. 10.1128/MCB.02190-06 PubMed 17296725
