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pcDNA3.1-KozATG-dL5-2XG4S-mCer3
(Plasmid #73207)

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This material is available to academics and nonprofits only.
Item Catalog # Description Quantity Price (USD)
Plasmid 73207 Standard format: Plasmid sent in bacteria as agar stab 1 $94
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DNA 73207-D50 50 μg of DNA in Tris buffer $495
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DNA 73207-D100 100 μg of DNA in Tris buffer $585
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Backbone

  • Vector backbone
    pcDNA3.1
  • Backbone manufacturer
    Invitrogen
  • Backbone size w/o insert (bp) 5382
  • Total vector size (bp) 6921
  • Modifications to backbone
    The NdeI site was removed from the CMV promoter
  • 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
    KozATG-dL5-2XG4S-mCer3
  • Alt name
    cyto
  • Species
    H. sapiens (human), Synthetic
  • Insert Size (bp)
    1539
  • Mutation
    The dL5** FAP is E50D, L89S, also known as E52D/L91S and NP138
  • Promoter CMV
  • Tag / Fusion Protein
    • The FAP and mCerulean3 are fused with 2 copies of a G4S linker

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

  • A portion of this plasmid was derived from a plasmid made by
    The mCerulean3 was 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/

Details of mCerulean3 can be found in:
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. PLoS One 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.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 50 μg
  • Concentration 1 μg/μL
  • Pricing $495 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

Purpose

Ready-to-use, high-purity DNA prep. DNA aliquots are suitable for use in mammalian cell transfections and other molecular biology applications.

Delivery

  • Amount 100 μg
  • Concentration 1 μg/μL
  • Pricing $585 USD
  • Storage 4 ℃ (short-term) or -20 ℃ (long-term)

Terms and Licenses

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

Quality Control

Every DNA prep is verified by next-generation sequencing, confirming plasmid identity, sequence integrity, and absence of DNA contaminants. Concentration is confirmed to be within ±2.5% of the stated concentration.

Our production process has been validated to consistently yield endotoxin levels below 1.0 EU/μg. Individual lots are not tested for endotoxin.

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-KozATG-dL5-2XG4S-mCer3 was a gift from Marcel Bruchez (Addgene plasmid # 73207 ; http://n2t.net/addgene:73207 ; RRID:Addgene_73207)

  • 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
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