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Depositing Lab
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Depositing OrganizationWashington University in Saint Louis
Located in United States of America -
Publication
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Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 20185 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 20185-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 20185-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonep3XFLAG-CMV-7
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Backbone manufacturerSigma
- Backbone size w/o insert (bp) 4700
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Vector typeMammalian Expression
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 nameNotch 3 Intracellular Domain
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Alt nameNICD3
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Alt nameNotch 3
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SpeciesM. musculus (mouse)
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Insert Size (bp)1962
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MutationContains murine Notch 3 Intracellular Domain (Met-1664 to Ala-2318)
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GenBank IDNM_008716.2 NP_032742.1
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Entrez GeneNotch3 (a.k.a. N3, hpbk)
- Promoter CMV
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Tag
/ Fusion Protein
- 3X Flag (N terminal on backbone)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site Xba1 (not destroyed)
- 3′ cloning site BamH1 (not destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer hGH-pA-R
- (Common Sequencing Primers)
Resource Information
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A portion of this plasmid was derived from a plasmid made byNotch 3 plasmid from Dr. Urban Lendahl (Karolinska Institutet, Stockholm, Sweden).
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
All NICD plasmids were cloned into the p3XFLAG-CMV-7(TM) expression vector (Sigma). NICD3 was cloned into XbaI/BamHI sites. NICD3 (Met-1664 to Ala-2318 of full-length mouse N3) was amplified from the N3 plasmid (gift from Dr. U. Lendahl) with 5XbaN3 (5'-GCTCTAGAATGGTTGCCAGGCGAAAGCGAG-3') and 3BamN3
(5'-GCGGATCCCTAGGCCATCACCTGCCTCTTGGG-3') primers.
DNA (Catalog # 20185-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 # 20185-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:
3XFlagNICD3 was a gift from Raphael Kopan (Addgene plasmid # 20185 ; http://n2t.net/addgene:20185 ; RRID:Addgene_20185) -
For your References section:
Target selectivity of vertebrate notch proteins. Collaboration between discrete domains and CSL-binding site architecture determines activation probability. Ong CT, Cheng HT, Chang LW, Ohtsuka T, Kageyama R, Stormo GD, Kopan R. J Biol Chem. 2006 Feb 24. 281(8):5106-19. 10.1074/jbc.M506108200 PubMed 16365048