pLPS-hAkt1-gfp
(Plasmid #49388)

• Purpose: mammalian expression of human Akt1 fused to EGFP
• Depositing Lab: Randall McKinnon
• Publication: McKinnon Lab plasmids (unpublished)

Backbone

• Vector backbone: pLPS-3'EGFP
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4900
• Vector type: Mammalian Expression, Cre/Lox ; acceptor vector
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Chloramphenicol and Kanamycin, 25 & 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: Akt1
• Alt name: v-akt murine thymoma viral oncogene homolog 1
• Species: H. sapiens (human)
• Insert Size (bp): 1487
• Mutation: tga to cca at nt 1725 with respect to GenBank ID BC000479
• GenBank ID: BC000479
• Entrez Gene: AKT1 (a.k.a. AKT, PKB, PKB-ALPHA, PRKBA, RAC, RAC-ALPHA)
• Promoter: CMV
• Tag / Fusion Protein:
  • EGFP (C terminal on backbone)

Cloning Information

• Cloning method: Unknown
• 5′ sequencing primer: CMV-F
• 3′ sequencing primer: EGFP-N

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

hAkt1 was transferred from the donor vector (pDNR-hAkt1, Addgene plasmid #49387) to the acceptor vector using Cre-mediated recombination at the LoxP sites.
When the recombinant construct is expressed in mammalian cells, EGFP is expressed from the immediate early promoter of cytomegalovirus (pCMV IE) as a 3' fusion to Akt1, through intron splicing.