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PurposeGFP reporter protein fused to three GSK3 phosphorylation sites
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Depositing Lab
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Depositing OrganizationUniversity of California, Los Angeles (UCLA)
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 | 29689 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 29689-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 29689-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepCS2
- Backbone size w/o insert (bp) 4096
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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 nameGFP glycogen synthase kinase 3 recognition site primed by MAPK
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Alt nameGFP fused to 3x GSK3 phosphorylation sites
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Insert Size (bp)121
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Mutationrecognition sites only, and E3 ligase site
- Promoter SP6
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Tags
/ Fusion Proteins
- Flag-EGFP (N terminal on backbone)
- Flag (C terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site EcoR1 (not destroyed)
- 3′ cloning site Xho1 (not destroyed)
- 5′ sequencing primer SP6
- 3′ 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
This artificial GFP-GSK3-MAPK construct was generated by adding to the C terminus of GFP synthetic oligonucleotides encoding for AAASAAASAAASAPASPAAAAAAAAPPAYDYKDDDDK. See Figure 6A of the associated publication for more details.
DNA (Catalog # 29689-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 # 29689-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:
pCS2 GFP-GSK3-MAPK was a gift from Edward De Robertis (Addgene plasmid # 29689 ; http://n2t.net/addgene:29689 ; RRID:Addgene_29689) -
For your References section:
Wnt signaling requires sequestration of glycogen synthase kinase 3 inside multivesicular endosomes. Taelman VF, Dobrowolski R, Plouhinec JL, Fuentealba LC, Vorwald PP, Gumper I, Sabatini DD, De Robertis EM. Cell. 2010 Dec 23. 143(7):1136-48. 10.1016/j.cell.2010.11.034 PubMed 21183076