Plasmid 14695: pRK5 BARK1 minigene

• Gene/insert name: BARK1 minigene
• Alt name: beta ARK1
• Alt name: beta adrenergic receptor kinase
• Alt name: ADRBK1
• Species: B. taurus (bovine)
• Entrez Gene: ADRBK1 (BOS_24950, GRK2)
  
• Fusion protein or tag: 3'UTR of human b-globin
• Terminal: C terminal on insert
• Mutation: BARK1 amino acids 495-689
• Vector backbone: pRK5
  (Search Vector Database)
• Vector type: Mammalian Expression
• Backbone size w/o insert: 4700
• Cloning site 5': EcoRI
• Site destroyed during cloning: No
• Cloning site 3': XbaI
• Site destroyed during cloning: No
• 5' sequencing primer: Sp6
• Bacterial resistance(s) Ampicillin
• Growth strain(s) DH5alpha
• Growth temperature (℃): 37
• High or low copy: High Copy
• Sequence: View sequences (1)
• Map: View map
• Principal Investigator: Robert Lefkowitz
• Terms and Licenses: MTA

Comments: 

The cDNA encoding bovine BARK1 residues Gly495 to Leu689 was amplified using PCR to construct a minigene casette with newly engineered 5' and 3' ends (see Author's map). The 5' end contained an EcoRI site for subcloning, followed by the kozak sequence 5'-GCCGCCACCATGG-3'. A bclI site was inserted at the 3' end immediately following the stop codon. The final BARK1 minigene construct contains the 3'UTR of the human beta-globin gene and a polyadenylation signal.

Article: Cellular expression of the carboxyl terminus of a G protein-coupled receptor kinase attenuates G beta gamma-mediated signaling. Koch et al (J Biol Chem. 1994 Feb 25. 269(8):6193-7. PubMed)