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Depositing Lab
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Publication
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Sequence Information
Full plasmid sequence is not available for this item.
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 10817 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
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.
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