pcDNA3.1-kappa-myc-dL5-2xG4S-TMst
(Plasmid #73206)

• Purpose: Expresses myc-dL5(E52D)-TM (PDGFR derived) on the surface of mammalian cells, with an Igk-leader. (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): 5388
• Total vector size (bp): 6459
• Modifications to backbone: The NdeI site was removed from the CMV promoter. The XhoI in the vector was destroyed. Another was introduced with the insert.
• 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: kappa-myc-dL5-2XG4S-TM
• Alt name: dL5-TM
• Alt name: Surface
• Species: H. sapiens (human)
• Insert Size (bp): 1071
• 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 is 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: SalI/XhoI but another XhoI introduced with the insert (destroyed during cloning)
• 5′ sequencing primer: CMVF
• 3′ sequencing primer: BGHR

Resource Information

• Supplemental Documents:
  • pcDNA3.1-kappa-myc-dL5-2XG4S-TMst
• Article Citing this Plasmid:
  • 1 Reference

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/

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.