pRK5 BARK1 minigene
(Plasmid #14695)

• Depositing Lab: Robert Lefkowitz
• Publication: Koch et al J Biol Chem. 1994 Feb 25. 269(8):6193-7.

Backbone

• Vector backbone: pRK5
• Backbone size w/o insert (bp): 4700
• Vector type: Mammalian Expression

Growth in Bacteria

• Bacterial Resistance(s): Ampicillin, 100 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: BARK1 minigene
• Alt name: beta ARK1
• Alt name: beta adrenergic receptor kinase
• Alt name: ADRBK1
• Species: B. taurus (bovine)
• Mutation: BARK1 amino acids 495-689
• Entrez Gene: GRK2 (a.k.a. ADRBK1)
• Tag / Fusion Protein:
  • 3'UTR of human b-globin (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: EcoRI (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: Sp6

Resource Information

• Articles Citing this Plasmid:
  • 3 References

Terms and Licenses

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

Depositor 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.