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Depositing Lab
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Depositing OrganizationChildren's Hospital of Eastern Ontario Research Institute
Located in Canada -
Publication
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Sequence Information
Full plasmid sequence is not available for this item.
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 20210 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 20210-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 20210-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA3
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5400
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Modifications to backboneThe expression plasmid pMC was constructed by inserting a synthetic polylinker (5′-AGC TTG CGG CCG CTA GCC TCG AGG GAT CCT CTA GAA TGC A-3′) into a pcDNA3 vector (Invitrogen) followed by insertion of the chloramphenicol acetyl transferase (CAT) coding sequence. The CAT insert was generated by PCR amplification using pCAT as a template.
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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 nameTNFa 3' UTR
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SpeciesH. sapiens (human)
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Insert Size (bp)808
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Entrez GeneTNF (a.k.a. DIF, IMD127, TNF-alpha, TNFA, TNFSF2, TNLG1F)
- Promoter CMV
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site XbaI (not destroyed)
- 5′ sequencing primer CAT-F (5'-GTTTGTGATGGCTTCCATGTC-3')
- (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
The 3′UTR was generated by PCR amplification from a liver cDNA library (Invitrogen). and the expression plasmids pMC-TNFα was constructed by inserting the 3′UTR of TNFα downstream of CAT in the pMC construct.
DNA (Catalog # 20210-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 # 20210-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:
pMC-TNFalpha was a gift from Martin Holcik (Addgene plasmid # 20210 ; http://n2t.net/addgene:20210 ; RRID:Addgene_20210) -
For your References section:
hnRNP A1 regulates UV-induced NF-kappaB signalling through destabilization of cIAP1 mRNA. Zhao TT, Graber TE, Jordan LE, Cloutier M, Lewis SM, Goulet I, Cote J, Holcik M. Cell Death Differ. 2009 Feb . 16(2):244-52. 10.1038/cdd.2008.146 PubMed 18846111