pcDNA C12 ecDHFR DD YFP HA
(Plasmid #192815)

• Purpose: Expresses an N-terminal enhanced destabilizing domain (DD) version of E. coli DHFR with higher basal turnover in mammalian systems. Contains missense mutations W74R/T113S/E120D/Q146L.
• Depositing Lab: John Hulleman
• Publication: Nakahara et al ACS Chem Biol. 2022 Sep 19. doi: 10.1021/acschembio.2c00518.

Backbone

• Vector backbone: pcDNA DEST40
• Backbone manufacturer: Thermo Fisher
• Backbone size w/o insert (bp): 5483
• Total vector size (bp): 6716
• 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
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: E. coli dihydrofolate reductase
• Alt name: ecDHFR
• Alt name: folA
• Species: E. coli
• Insert Size (bp): 1233
• Mutation: W74R/T113S/E120D/Q146L
• GenBank ID: J01609.1
• Promoter: CMV
• Tags / Fusion Proteins:
  • enhanced yellow fluorescent protein (C terminal on insert)
  • hemagglutinin (C terminal on insert)

Cloning Information

• Cloning method: Gibson Cloning
• 5′ sequencing primer: CMV For

Terms and Licenses

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