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Purposemammalian expression of mRFP1-tagged mouse Smad2
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Depositing Lab
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Depositing OrganizationUniversity of Virginia
Located in United States of America -
Sequence Information
Full plasmid sequence is not available for this item.
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 58491 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 58491-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 58491-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepBabe N-terminal RFP neo
- Backbone size w/o insert (bp) 5729
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Modifications to backboneThis construct is a variant of pBabe neo that expresses mRFP1 as an N-terminal fusion. A new XhoI site was inserted immediately upstream of the BamHI site by PCR during cloning. TO EXPRESS AN N-mRFP1 FUSION, GENES MUST HAVE A BAMHI- OR XHO1-COMPATIBLE 5' STICKY END.
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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)Stbl3
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert nameSmad2
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Alt nameSMAD family member 2
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SpeciesM. musculus (mouse)
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Insert Size (bp)1414
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GenBank IDNM_010754
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Entrez GeneSmad2 (a.k.a. 7120426M23Rik, Madh2, Madr2, Smad-2, mMad2)
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Tag
/ Fusion Protein
- mRFP1 (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (destroyed during cloning)
- 3′ cloning site SalI (not destroyed)
- 5′ sequencing primer pBABE 5'
- 3′ sequencing primer pBABE 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
pBabe RFP1_Smad2 neo was constructed by PCR cloning from plasmid templates (Open Biosystems) into the retroviral vector pBabe N-terminal RFP neo. The PCR product was gel purified, digested with BclI and SalI, and then ligated into pBabe N-terminal RFP neo, which had been digested with BamHI and SalI and then dephosphorylated.
DNA (Catalog # 58491-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 # 58491-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:
pBabe RFP1-Smad2 neo was a gift from Kevin Janes (Addgene plasmid # 58491 ; http://n2t.net/addgene:58491 ; RRID:Addgene_58491) -
For your References section:
A time- and matrix-dependent TGFBR3-JUND-KRT5 regulatory circuit in single breast epithelial cells and basal-like premalignancies. Wang CC, Bajikar SS, Jamal L, Atkins KA, Janes KA. Nat Cell Biol. 2014 Apr;16(4):345-56. doi: 10.1038/ncb2930. Epub 2014 Mar 23. 10.1038/ncb2930 PubMed 24658685