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Depositing Lab
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Depositing OrganizationBeth Israel Deaconess Medical Center
Located in the United States of America -
Publication
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Sequence Information
Full plasmid sequence is not available for this item.
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | |
|---|---|---|---|---|---|
| Plasmid | 10817 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $94 | |
| DNA | 10817-D50 | 50 μg of DNA in Tris buffer | $495 | ||
| DNA | 10817-D100 | 100 μg of DNA in Tris buffer | $585 | ||
Backbone
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Vector backbonepcDNA3
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5446
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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 namePKD
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SpeciesH. sapiens (human)
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Insert Size (bp)2800
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MutationS910A--active; L520P
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Entrez GenePRKD1 (a.k.a. CHDED, PKC-MU, PKCM, PKD, PRKCM)
- Promoter CMV
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Tag
/ Fusion Protein
- HA (N terminal on insert)
Cloning Information
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (not destroyed)
- 3′ cloning site XhoI (not destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer BGH-rev
- (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
The HA tag on this plasmid contains a point mutation that renders it unrecognizable by the 3F10 HA antibody. It will work fine with the 12CA5 HA antibody.
The PKD insert in this plasmid contains a L520P mutation when compared to GenBank reference sequence NP_002733.1. Although the effect of this mutation is unknown, this is the exact plasmid used in the associated publication and it should function as described.
Note that the S916A mutation is S910A when numbered according to GenBank reference sequence NP_002733.1.
DNA (Catalog # 10817-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 # 10817-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:
HA.PKD.S916A was a gift from Alex Toker (Addgene plasmid # 10817 ; http://n2t.net/addgene:10817 ; RRID:Addgene_10817) -
For your References section:
Activation loop phosphorylation controls protein kinase D-dependent activation of nuclear factor kappaB. Storz P, Doppler H, Toker A. Mol Pharmacol. 2004 Oct . 66(4):870-9. 10.1124/mol.104.000687 PubMed 15226414