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pBRα-β flap (831-1057)
(Plasmid #53734)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 53734 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    NEB5alpha F'LacIQ
  • Growth instructions
    We recommend using a lacIQ strain, such as NEB5alpha F'LacIQ, in order to ensure that the expression of potentially toxic fusion proteins is kept repressed.
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    alpha NTD-Beta flap fusion
  • Alt name
    rpoA-rpoB fusion
  • Promoter lpp/lacUV5 (tandem promoter)

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NotI (not destroyed)
  • 3′ cloning site BamHI (not destroyed)
  • 5′ sequencing primer pBRa_F (approx 150bp upstream of NotI site) 5’ gaacagcgtaccgacctggac 3’
  • (Common Sequencing Primers)

Resource Information

Depositor Comments

Note-When cloning into this plasmid, the 5' end of the insert must provide an extra nucleotide to maintain the reading frame. Additionally, a stop codon must be inserted 5' to the BamHI site. Please see supplemental document for more details.

This plasmid is used in conjuction with the other Ann Hochschild Bacterial 2-hybrid system plasmids (Addgene plasmids 53730, 53731, 53732, 53733, and 53734) and must be used in the reporter strain, FW102 OL2–62 (Addgene item 53735). Please see the associated article for links to all of these plasmids:
http://www.addgene.org/browse/article/8393/

The following articles can provide more information on using the bacterial 2-hybrid system:
Dove, et al. 1997 PMID 9121589
Dove and Hochschild 1998 PMID 9499408
Deaconescu, et al. 2006 PMID 16469698
Deighan, et al. 2008 PMID 18832144
Kuznedelov, et al. 2002 PMID 11823642

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:

    pBRα-β flap (831-1057) was a gift from Ann Hochschild (Addgene plasmid # 53734)
  • For your References section:

    Activation of prokaryotic transcription through arbitrary protein-protein contacts. Dove SL, Joung JK, Hochschild A. Nature. 1997 Apr 10;386(6625):627-30. 10.1038/386627a0 PubMed 9121589