Part:BBa_K4129102

FunsTF04

FunsTF04 is a synthetic transcription factor (sTF). FunsTF04 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor. FunsTF04 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: Hmox1, transactivation domain; B112 and the nuclear localization signal (NLS) SV40. The linker between LexA and Hmox1 was a longer version (Ottoz et. al (2014) compared to sBAD (Castaño-Cerezo et. al (2020)). LexA is a repressor that regulates the SOS response in E. coli (Radman. 1975). LexA binds to a specific DNA motif, lexO (Erill. et al (2003)), and it is the DNA binding domain that interacts with LexO that is used in FunsTF04. Hmox1 is the human heme oxygenase 1, which can cleave heme. This function is not of our interest, but it was shown that furfural binds to hmox1. The transactivation domain B112 is from E. coli, which were experimentally proven to initiate transcription of a synthetic promoter in S. cerevisiae (Ottoz et. al (2014)). The NLS SV40 is a small peptide sequence of PKKKRKV that enables transport og the protein to the nucleus (Garcia-Bustos et. al (1991)).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 945
  Illegal BamHI site found at 607
  Illegal BamHI site found at 1604
  Illegal XhoI site found at 800
  Illegal XhoI site found at 1237
  Illegal XhoI site found at 1753
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]