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, MGC99656, 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: 6600
• 5' sequencing primer: CMV Forward
• 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.

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)