Part:BBa_K4825021
pNOTAra1
pNOTAra1 is a synthetic repressible promoter designed for automation for yeast, in which the level of the co-repressor arabinose regulates the initiation of the transcription. AraC functions as a homodimer. In the absence of arabinose, the promoter is accessible. In the presence of arabinose, two subunits of AraC bind to adjacent Inv Aral1 and Inv Ara2 separated by four bp and the two subunits are connected through the dimerisation domain. In our project, this part is utilized to construct a plasmid with BAX gene and control its transcription by repressing it with the presence of arabinose in S.cerevisiae and prevent the initiation of kill switch.
Usage and Biology
Ara promoter with L-arabinose operon and its downstream araBAD gene regulates the metabolisation of arabinose in E.coli. In natural E.coli cell system, the regulatory protein of Ara operon, AraC, acts positively to stimulate transcription in the presence of arabinose and acts negatively in the absence of arabinose to repress transcription initiation. In the presence of high level of arabinose and low level of glucose, the complex of cAMP and catabolite activator protein(CAP) together with dimeric AraC protein with two subunits binding to AraI 1 and Ara 2 elements respectively, stimulate the initiation. In the absence of arabinose, the dimeric AraC with two subunits binding to AraI1 and AraO2 elements respectively represses the transcription. AraI with two half sites are selected to design the synthetic pNOTara1 promoter in this part.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 1
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1
- 1000COMPATIBLE WITH RFC[1000]
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