pCDNA3.1 AT1AR-TSTS/4A
(Plasmid #45634)

• Depositing Lab: Robert Lefkowitz
• Depositing Organization: Duke University
  Located in United States of America
• 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): 6254
• 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-TSTS/A
• Alt name: AT1AR
• Alt name: Agtr1
• Alt name: angiotensin II receptor, type 1
• Species: R. norvegicus (rat)
• Insert Size (bp): 1254
• Mutation: Thr332, Ser335, Thr336, and Ser338 all changed to Alanine
• 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 TSTS/A receptor was:
5' p-CTCAAGCCTGTCTGCGAAAATGGCCGCTTGCTTACCGGCCTTC 3'.

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

DNA (Catalog # 45634-D50)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 50 μg
• Concentration: 1 μg/μL
• Pricing: $495 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

DNA (Catalog # 45634-D100)

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

• Amount: 100 μg
• Concentration: 1 μg/μL
• Pricing: $585 USD
• Storage: 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.