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MBP_TS-B1AR_Trx_PreSc_NdeI_AgeI
(Plasmid #247768)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 247768 Standard format: Plasmid sent in bacteria as agar stab 1 $89
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Backbone

  • Vector backbone
    pRG/III-hs-MBP
  • Backbone manufacturer
    Reinhard Grisshammer and Richard Henderson, Addgene plasmid #121676
  • Modifications to backbone
    Thrombin cleavage sites were replaced with the more selective HRV 3C protease recognition sites to improve specificity. Each cleavage site was flanked on both sides by flexible (Gly-Ser)5 linkers to minimize structural interference with the fusion proteins or target protein. To facilitate future replacement of the β1AR gene with other GPCRs or transmembrane proteins, we introduced NdeI and AgeI restriction sites upstream and downstream of the β1AR coding sequence, respectively.
  • Vector type
    Bacterial Expression

Growth in Bacteria

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

Gene/Insert

  • Gene/Insert name
    Thermostable Turkey B1-adrenergic receptor (TS-B1AR)
  • Species
    Meleagris gallopavo
  • Promoter lac promoter
  • Tag / Fusion Protein
    • MBP–(GS)5–HRV 3C site–(GS)5–NdeI site_ B1AR_AgeI site–(GS)5 linker–HRV 3C cleavage site–(GS)5–TrxA–10 x His tag

Cloning Information

  • Cloning method Ligation Independent Cloning
  • 5′ sequencing primer aac atc ccg cag atg tcc gct ttc tgg tat gcc gtg c
  • 3′ sequencing primer tcc tgg tag cac ttg agc gcc tga ggg tcc tc
  • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    The original plasmid pRG/III-hs-MBP was kindly provided by R. Grisshammer (NIH).

Terms and Licenses

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

Trademarks:

  • Zeocin® is an InvivoGen trademark.
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:

    MBP_TS-B1AR_Trx_PreSc_NdeI_AgeI was a gift from Stephan Grzesiek (Addgene plasmid # 247768 ; http://n2t.net/addgene:247768 ; RRID:Addgene_247768)

  • For your References section:

    Efficient production of a functional G protein-coupled receptor in E. coli for structural studies. Abiko LA, Rogowski M, Gautier A, Schertler G, Grzesiek S. J Biomol NMR. 2021 Jan;75(1):25-38. doi: 10.1007/s10858-020-00354-6. Epub 2021 Jan 27. 10.1007/s10858-020-00354-6 PubMed 33501610
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