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Depositing Lab
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Depositing OrganizationMemorial Sloan-Kettering Cancer Center
Located in the United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 14959 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 14959-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 14959-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA3.1(+)
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Backbone manufacturerInvitrogen
- 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 nameSmad4
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SpeciesH. sapiens (human)
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Insert Size (bp)1700
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Entrez GeneSMAD4 (a.k.a. DPC4, JIP, MADH4, MYHRS)
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Tag
/ Fusion Protein
- Flag (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site XhoI / SalI (destroyed during cloning)
- 5′ sequencing primer CMV Forward
- (Common Sequencing Primers)
Resource Information
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Addgene Notes
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
Site directed mutagenesis in the coding sequence of Tyr162 (from TAT to TAC) and Val163 (from GTG to GTT). The mutated sequence encodes the same amino acids. See author's map for more information.
DNA (Catalog # 14959-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 # 14959-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:
pcDNA Flag-Smad4M was a gift from Joan Massague (Addgene plasmid # 14959 ; http://n2t.net/addgene:14959 ; RRID:Addgene_14959) -
For your References section:
Breast cancer bone metastasis mediated by the Smad tumor suppressor pathway. Kang Y, He W, Tulley S, Gupta GP, Serganova I, Chen CR, Manova-Todorova K, Blasberg R, Gerald WL, Massague J. Proc Natl Acad Sci U S A. 2005 Sep 27. 102(39):13909-14. 10.1073/pnas.0506517102 PubMed 16172383