pcDNA3.1-kappa-myc-dL5-2XG4S-mCer3-KDEL
(Plasmid
#73209)
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PurposeExpresses myc-dL5(E52D)-mCer3 fusion protein in endoplasmic reticulum of mammalian cells with an Igk-leader. (MBIC5, dL5**, FAP)
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Depositing Lab
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Sequence Information
Ordering
Item | Catalog # | Description | Quantity | Price (USD) | |
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Plasmid | 73209 | Standard format: Plasmid sent in bacteria as agar stab | 1 | $85 |
Backbone
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Vector backbonepcDNA3.1
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Backbone manufacturerInvitrogen
- Backbone size w/o insert (bp) 5393
- Total vector size (bp) 7062
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Modifications to backboneNdeI site was removed from the CMV promoter. XhoI site duplicated for insertion of KDEL coding sequence and STOP codon.
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Vector typeMammalian Expression
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Selectable markersNeomycin (select with G418)
Growth in Bacteria
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Bacterial Resistance(s)Ampicillin, 100 μg/mL
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Growth Temperature37°C
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Growth Strain(s)DH5alpha
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Growth instructionsNo special instructions.
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Copy numberHigh Copy
Gene/Insert
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Gene/Insert namekappa-myc-dL5-2XG4S-mCer3-KDEL
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Alt nameER
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SpeciesH. sapiens (human), Synthetic
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Insert Size (bp)1674
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MutationThe dL5** FAP is E50D, L89S, also known as E52D/L91S and NP138
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Entrez GeneMYC (a.k.a. MRTL, MYCC, bHLHe39, c-Myc)
- Promoter CMV
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Tags
/ Fusion Proteins
- The FAP and mCerulean3 are fused with 2 copies of a G4S linker
- myc 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
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Supplemental Documents
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A portion of this plasmid was derived from a plasmid made byThe mCerulean3 is from MA Rizzo, Department of Physiology, University of Maryland School of Medicine, Baltimore, Maryland, United States of America
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Article Citing this Plasmid
Terms and Licenses
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Academic/Nonprofit Terms
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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.
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.
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.
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For your Materials & Methods section:
pcDNA3.1-kappa-myc-dL5-2XG4S-mCer3-KDEL was a gift from Marcel Bruchez (Addgene plasmid # 73209 ; http://n2t.net/addgene:73209 ; RRID:Addgene_73209) -
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