Difference between revisions of "Part:BBa K4129110"
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FunsTF61 is a synthetic transcription factor (sTF).  FunsTF61 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor.   
 
FunsTF61 is a synthetic transcription factor (sTF).  FunsTF61 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor.   
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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===Functional Parameters===
 
===Functional Parameters===
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Revision as of 21:29, 10 October 2022

The fungal synthetic transcription factor, FunsTF61 (LexA-LL-HbaR7-VP16-SV40)

FunsTF61 is a synthetic transcription factor (sTF). FunsTF61 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor. FunsTF61 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR7, transactivation domain; VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR7 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 FunsTF61. HbaR is a transcriptional factor from Rhodopseudomonas palustris that initiates transcription in the presence of benzoic acid or in the presence of benzoic acid derivatives (Egland. Et al (2000) (Castaño-Cerezo et. al (2020)). We created 16 mutants of HbaR and FunsTF61 carried mutant 7 of HbaR, which had the following mutations: A45V, L69A, G71K, E77M, Y96A, L97F, N99T, A100V and V145M. Viral Protein 16 (VP16) from herpes simplex virus type 1 is a transcription factor that uses a transactivation domain to recruit the RNA polymerase II.The NLS SV40 is a small peptide sequence of PKKKRKV that enables transport of the protein to the nucleus (Garcia-Bustos et. al (1991)).

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 673
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 607
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 765
  • 1000
    COMPATIBLE WITH RFC[1000]