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Addgene

pcDNA3.1-NLS-myc-dL5-2xG4S-mCer3
(Plasmid #73205)

Ordering

This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 73205 Standard format: Plasmid sent in bacteria as agar stab 1 $85

Backbone

  • Vector backbone
    pcDNA3.1
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5382
  • Total vector size (bp) 7011
  • Modifications to backbone
    NdeI site was destroyed by adding tata into the CMV promoter at 487-490
  • Vector type
    Mammalian Expression
  • Selectable markers
    Neomycin (select with G418)

Growth in Bacteria

  • Bacterial Resistance(s)
    Ampicillin, 100 μg/mL
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Growth instructions
    No special instructions.
  • Copy number
    High Copy

Gene/Insert

  • Gene/Insert name
    NLS-myc-dL5-2XG4S-mCer3
  • Alt name
    NLS
  • Species
    H. sapiens (human), Synthetic
  • Insert Size (bp)
    1623
  • Mutation
    The dL5** FAP is E50D, L89S, also known as E52D/L91S and NP138
  • Entrez Gene
    MYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
  • Promoter CMV
  • Tags / Fusion Proteins
    • The FAP and mCerulean3 are fused with 2 copies of a G4S linker
    • myc epitope 1020-1049

Cloning Information

  • Cloning method Restriction Enzyme
  • 5′ cloning site BamHI (not destroyed)
  • 3′ cloning site XhoI (not destroyed)
  • 5′ sequencing primer CMVF
  • 3′ sequencing primer BGHR
  • (Common Sequencing Primers)

Resource Information

  • Supplemental Documents
  • A portion of this plasmid was derived from a plasmid made by
    The coding sequence for mCerulean3 was a gift from MA Rizzo, Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America

Terms and Licenses

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

Depositor Comments

Please visit the Molecular Biosensor and Imaging Center website for current protocols and information about dyes to be used in combination with the proteins expressed from this plasmid.

http://pathways.mbic.cmu.edu/

For information concerning mCerulean3 please refer to:
Markwardt, M. L., Kremers, G. J., Kraft, C. A., Ray, K., Cranfill, P. J., Wilson, K. A., Day, R. N., Wachter, R. M., Davidson, M. W., and Rizzo, M. A. (2011) An Improved Cerulean Fluorescent Protein with Enhanced Brightness and Reduced Reversible Photoswitching. PLoSOne 6, e17896.

For more information about this plasmid, please see the associated publication (PMID: 25650487) as well as He et al. Nat Methods. 2016 Jan 25. doi: 10.1038/nmeth.3735. [Epub ahead of print] PMID: 26808669.

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-NLS-myc-dL5-2xG4S-mCer3 was a gift from Marcel Bruchez (Addgene plasmid # 73205 ; http://n2t.net/addgene:73205 ; RRID:Addgene_73205)
  • For your References section:

    Rapid, specific, no-wash, far-red fluorogen activation in subcellular compartments by targeted fluorogen activating proteins. Telmer CA, Verma R, Teng H, Andreko S, Law L, Bruchez MP. ACS Chem Biol. 2015 May 15;10(5):1239-46. doi: 10.1021/cb500957k. Epub 2015 Feb 16. 10.1021/cb500957k PubMed 25650487