pCDNA3.1 AT1AR-324delta
(Plasmid #45635)

• Depositing Lab: Robert Lefkowitz
• Publication: Wei et al J Biol Chem. 2004 Nov 12;279(46):48255-61. Epub 2004 Sep 7.

Backbone

• Vector backbone: pcDNA3.1/Zeo
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5000
• Total vector size (bp): 6250
• Vector type: Mammalian Expression
• Selectable markers: Zeocin

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: AT1AR-324delta
• Alt name: AT1AR
• Alt name: Agtr1
• Alt name: angiotensin II receptor, type 1
• Species: R. norvegicus (rat)
• Mutation: added stop codon at 324 aa
• Entrez Gene: Agtr1a (a.k.a. AT1, AT1A, AT1R, Agtr1)
• Promoter: CMV
• Tag / Fusion Protein:
  • HA (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: HindIII (not destroyed)
• 3′ cloning site: XbaI (not destroyed)
• 5′ sequencing primer: CMV-F; T7
• 3′ sequencing primer: BGH-rev

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The template pcDNA3.1 expression plasmid encoding hemagglutinin (HA) epitope-tagged AT1A receptors was provided by M. G. Caron (Duke University).

Terms and Licenses

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

Depositor Comments

Mutant AT1A receptors were generated by mutagenesis PCR using pcDNA3.1-HA-AT1A receptor (provided by M. G. Caron, Duke University) as a template and a QuikChange multisite-directed mutagenesis kit (Stratagene) according to the manufacturer’s instructions. The primer for the 324delta receptor was:
5' p-CCCCCAAAGGCCTAGCTCCCACTCAAGCC 3'.

N4D mutation introduced to prevent glycosylation at this consensus site and steric hindrance of antibody binding.