- Backbone size w/o insert (bp) 4900
Vector typeMammalian Expression
Growth in Bacteria
Copy numberHigh Copy
Full plasmid sequence is available only if provided by the depositing laboratory.
SpeciesH. sapiens (human)
Insert Size (bp)2000
MutationCatalytically inactive Pak1: K299R
Entrez GenePAK1 (a.k.a. PAKalpha)
/ Fusion Protein
- Myc (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site SalI (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer CTCGTTTAGTGAACCGTCAG
- 3′ sequencing primer GGAACTTCCAAGGCCAGGA (Common Sequencing Primers)
Please note that the reference sequence from the depositor does not have the K299R mutation. Addgene has sequenced this plasmid and verified this mutation.
There is also an additional mutation in this plasmid - L516I. The L516I mutation, or SNP, has been present from the very beginning (~1995). The crystal structure of Pak1 uses this clone, so it is present there too. The depositor does not think it affects function in any way.
Pak1 may contain a G401S mutation. Depositor states that this mutation should not affect plasmid function.
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:pCMV6M-Pak1 K299R was a gift from Jonathan Chernoff (Addgene plasmid # 12210)
For your References section:Human p21-activated kinase (Pak1) regulates actin organization in mammalian cells. Sells MA, Knaus UG, Bagrodia S, Ambrose DM, Bokoch GM, Chernoff J. Curr Biol. 1997 Mar 1. 7(3):202-10. 10.1016/S0960-9822(97)70091-5 PubMed 9395435
Map generated by Addgene from plasmid data.