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

Full plasmid sequence is not available for this item.

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 45634 Standard format: Plasmid sent in bacteria as agar stab 1 $85

This material is available to academics and nonprofits only.

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
  • (Common Sequencing Primers)

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
  • Industry Terms
    • Not Available to Industry
Trademarks:
  • Zeocin® is an InvivoGen trademark.

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.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    pCDNA3.1 AT1AR-TSTS/4A was a gift from Robert Lefkowitz (Addgene plasmid # 45634 ; http://n2t.net/addgene:45634 ; RRID:Addgene_45634)
  • For your References section:

    Stable interaction between beta-arrestin 2 and angiotensin type 1A receptor is required for beta-arrestin 2-mediated activation of extracellular signal-regulated kinases 1 and 2. Wei H, Ahn S, Barnes WG, Lefkowitz RJ. J Biol Chem. 2004 Nov 12;279(46):48255-61. Epub 2004 Sep 7. 10.1074/jbc.M406205200 PubMed 15355986