Part:BBa_K4129111
The fungal synthetic transcription factor, FunsTF62 (LexA-LL-HbaR8-VP16-SV40)
FunsTF62 is a synthetic transcription factor (sTF). FunsTF62 should initiate the transcription through the 6xLexO minimal promoter. This sTF is the sensing part of the biosensor. FunsTF62 is a fusion protein consisting of the DNA-binding domain: lexA, ligand sensing domain: HbaR8, transactivation domain; VP16 and the nuclear localization signal (NLS) SV40. The linker between LexA and HbaR8 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 FunsTF62. 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 FunsTF62 carried mutant 8 of HbaR, which had the following mutations: A45V, L69A, G71K, E77A, A86G, E89G, A90G, A91P, Y96A, L97Y, A98S, N99T, A100V and V145Y. 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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 673
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 607
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 765
- 1000COMPATIBLE WITH RFC[1000]
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