pcDNA3.1-KozATG-myc-dL5-2XG4S-mCer3-SKL
(Plasmid #73210)

• Purpose: Expresses myc-dL5(E52D)-mCer3 fusion protein in peroxisomes of mammalian cells with a C-terminal PTS (SKL). (MBIC5, dL5**, FAP)
• Depositing Lab: Marcel Bruchez
• Publication: Telmer et al ACS Chem Biol. 2015 May 15;10(5):1239-46. doi: 10.1021/cb500957k. Epub 2015 Feb 16.

Backbone

• Vector backbone: pcDNA3.1
• Backbone manufacturer: Invitrogen
• Backbone size w/o insert (bp): 5397
• Total vector size (bp): 6957
• Modifications to backbone: The NdeI site was removed in 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-myc-dL5-2XG4S-mCer3-SKL
• Alt name: SKL
• Alt name: peroxisome
• Species: H. sapiens (human), Synthetic
• Insert Size (bp): 1560
• 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 951-980 (N terminal on insert)

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

Resource Information

• Supplemental Documents:
  • pcDNA3.1-myc-dL5-2XG4S-mCer3SKL
• A portion of this plasmid was derived from a plasmid made by: The mCerulean3 is from MA Rizzo, Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America

Terms and Licenses

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

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.