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Depositing Lab
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Depositing OrganizationDuke University
Located in United States of America -
Citations
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Sequence Information
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 32453 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 32453-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 32453-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepEGFP-N1
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Backbone manufacturerClontech
- Backbone size w/o insert (bp) 4700
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Modifications to backboneFLAG tag added into NheI site by cutting and inserting linker with FLAG tag and NheI overhangs. Second NheI site removed by site-directed mutagenesis.
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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)Kanamycin, 50 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Copy numberUnknown
Gene/Insert
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Gene/Insert nameCalcium/Calmodulin dependent protein kinase kinase beta
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Alt nameCaMKKbeta
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Alt nameCaMKK2
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SpeciesM. musculus (mouse)
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GenBank IDNM_145358.1
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Entrez GeneCamkk2 (a.k.a. 6330570N16Rik, AW061083, mKIAA0787)
- Promoter CMV
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Tags
/ Fusion Proteins
- GFP (C terminal on backbone)
- FLAG (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site NheI (not destroyed)
- 3′ cloning site SalI (not destroyed)
- 5′ sequencing primer CMV
- (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
A W374C mutation is also present in the pEGFP-N1-CaMKKbeta constructs that were used in this publication.
DNA (Catalog # 32453-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 # 32453-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:
pEGFP-N1-CaMKKbeta was a gift from Anthony Means (Addgene plasmid # 32453 ; http://n2t.net/addgene:32453 ; RRID:Addgene_32453) -
For your References section:
Ca2+/Calmodulin-dependent Protein Kinase Kinase {beta} Is Regulated by Multisite Phosphorylation. Green MF, Scott JW, Steel R, Oakhill JS, Kemp BE, Means AR. J Biol Chem. 2011 Aug 12;286(32):28066-79. Epub 2011 Jun 13. 10.1074/jbc.M111.251504 PubMed 21669867
