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Filamin-M-stFRET
(Plasmid #61105)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 61105 Standard format: Plasmid sent in bacteria as agar stab 1 $75

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pEYFP–C1
  • Backbone manufacturer
    Clontech
  • Backbone size w/o insert (bp) 4700
  • Total vector size (bp) 13552
  • Modifications to backbone
    Gene was subcloned into the pEYFP-C1 vector in which the yellow fluorescent protein (YFP) gene was deleted.
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Kanamycin
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Unknown

Gene/Insert

  • Gene/Insert name
    filamin-stFRET
  • Promoter CMV

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site NheI (unknown if destroyed)
  • 3′ cloning site Unknown (unknown if destroyed)
  • 5′ sequencing primer CMV Forward
  • 3′ sequencing primer SV40 poly A Reverse
  • (Common Sequencing Primers)

Resource Information

  • A portion of this plasmid was derived from a plasmid made by
    pEYFP–C1 Venus and pECFP–C1 Cerulean plasmids were generous gifts from David W. Piston. Cerulean gene was sub-cloned from pECFP-C1 and EcoRI and ApaI restriction enzyme sites were introduced into 5 prime and 3 prime of Cerulean DNA fragment. This DNA fragment was inserted into multiple cloning site of pEYFP-C1 Venus by EcoRI and ApaI digestion then ligation. The resulting vector has Venus followed closely by Cerulean and between them there are two restriction enzyme sites BglII and EcoRI which then were employed to insert the alpha-helix linker. The alpha-helix linker DNA was amplified by PCR and BglII and EcoRI sites were introduced into 5 prime and 3 prime ends. The final construct with alpha-helix connecting Venus and Cerulean was named stFRET and ready for eukaryotic expression.
  • Terms and Licenses
  • Industry Terms
    • Not Available to Industry
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:

    Filamin-M-stFRET was a gift from Fred Sachs (Addgene plasmid # 61105 ; http://n2t.net/addgene:61105 ; RRID:Addgene_61105)
  • For your References section:

    A fluorescence energy transfer-based mechanical stress sensor for specific proteins in situ. Meng F, Suchyna TM, Sachs F. FEBS J. 2008 Jun;275(12):3072-87. doi: 10.1111/j.1742-4658.2008.06461.x. Epub 2008 May 10. 10.1111/j.1742-4658.2008.06461.x PubMed 18479457