Depositor Sequences: None.
Addgene Sequences: Partial (4)
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||10809||Plasmid sent as bacteria in agar stab||1||$65|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 5446
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
SpeciesH. sapiens (human)
Insert Size (bp)2800
MutationKinase dead mutant: K612W (Lys 612 to Trp) (bp1834-1836 AAA -> TGG); Also Q66R, F139S and G877W
Entrez GenePRKD1 (a.k.a. CHDED, PKC-MU, PKCM, PKD, PRKCM)
- Promoter CMV
/ Fusion Proteins
- HA (N terminal on insert)
- Myc (inserted between amino acids 33&34 of PKD)
- 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)
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 Myc tag inserted between amino acids 33&34, Q66R, F139S and G877W mutations when compared to GenBank reference sequence NP_002733.1. Although the effect of these mutations is unknown, this is the exact plasmid used in the associated publication and it should function as described.
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.
For your Materials & Methods section:HA.PKD.K/W was a gift from Alex Toker (Addgene plasmid # 10809)
For your References section:Tyrosine phosphorylation of protein kinase D in the pleckstrin homology domain leads to activation. Storz P, Doppler H, Johannes FJ, Toker A. J Biol Chem. 2003 May 16. 278(20):17969-76. 10.1074/jbc.M213224200 PubMed 12637538
Map uploaded by the depositor.