Addgene - 902 pLNCX myr HA Akt1 K179M Plasmid Data

Plasmid 9006: 902 pLNCX myr HA Akt1 K179M

Gene/insert name:
Akt1 K179M
Alt name:
Akt1
Alt name:
Akt
Insert size:
1500
Species:
H. sapiens (human)
Entrez Gene:
AKT1 (AKT, CWS6, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA)
Fusion protein or tag:
myr
Terminal:
N terminal on insert
Fusion protein or tag:
HA
Terminal:
N terminal on insert
Mutation:
K179M. kinase dead.
Vector backbone:
pLNCX
(Search Vector Database)
Backbone manufacturer:
BD Biosciences
Vector type:
Mammalian Expression, Retroviral
Backbone size w/o insert (bp):
6600
5' sequencing primer:
CMV Forward List of Sequencing Primers
Bacterial resistance(s)
Ampicillin
Growth strain(s)
DH5alpha
Growth temperature (℃):
37
High or low copy:
High Copy
Selectable markers:
Neomycin
Sequence: View sequences (1)
Principal Investigator:
William Sellers
Terms and Licenses:
MTA

Comments:

An Akt-1 cDNA was amplified by reverse transcription-coupled PCR from total HeLa cell RNA and reamplified with a 5' primer containing a Kozak sequence and sequences encoding a hemagglutinin (HA) epitope and cloned into pLNCX to give pLNCX-HA-Akt. A double-stranded oligonucleotide encoding the src myristoylation sequence was inserted 5' of the HA tag to generate pLNCX-Myr-HA-Akt. K179M mutation generated using site-directed mutagenesis.

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.

Article: Regulation of G1 progression by the PTEN tumor suppressor protein is linked to inhibition of the phosphatidylinositol 3-kinase/Akt pathway. Ramaswamy et al (Proc Natl Acad Sci U S A. 1999 Mar 2. 96(5):2110-5. PubMed)

Please acknowledge the principal investigator and cite this article if you use this plasmid in a publication. Also, please include the text "Addgene plasmid 9006" in your Materials and Methods section.