Part:BBa_K5477024
pRAD27-AhR - receptor module
The pRAD27-AhR receptor module integrates the Aryl Hydrocarbon Receptor (AhR) BBa_K3793004 with the pRAD27 promoter BBa_K5477002, creating a system that is responsive to both environmental toxins and cellular stress related to DNA repair processes. The pRAD27 promoter is derived from the RAD27 gene, which is involved in DNA repair and replication in yeast, particularly in the processing of Okazaki fragments during DNA synthesis.
In this composite module, the pRAD27 promoter controls the expression of AhR, a ligand-activated transcription factor that senses and responds to environmental contaminants such as polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). When these toxins bind to AhR, the receptor translocates to the nucleus, where it dimerizes with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs) in the DNA, activating the transcription of detoxification genes, such as those coding for cytochrome P450 enzymes, which metabolize and neutralize harmful compounds.
The pRAD27-AhR module is used for our biosensor system for detecting environmental pollutants like PAHs and PCBs. Under the presence of these compounds, receptor modules including ARNT () and NCOA () will bind to XRE () and initiate the transcription of NanoLuc () in our reporter module (). The composite part was cloned using the method of USER-cloning into YCp-L. The YCp-L plasmid is a centromeric plasmid that carries a LEU2 marker for leucine selection in yeast. Centromeric plasmids like YCp-L are low-copy vectors, typically maintained at one to two copies per cell due to the presence of a CEN sequence. This composite part was used in our device:
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 1064
Illegal SpeI site found at 289
Illegal SpeI site found at 426 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1064
Illegal NheI site found at 869
Illegal SpeI site found at 289
Illegal SpeI site found at 426 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1064
Illegal BglII site found at 102
Illegal BglII site found at 1879 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 1064
Illegal SpeI site found at 289
Illegal SpeI site found at 426 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 1064
Illegal SpeI site found at 289
Illegal SpeI site found at 426 - 1000COMPATIBLE WITH RFC[1000]
References
1. Carambia, A., Schuran, F.A. The aryl hydrocarbon receptor in liver inflammation. Semin Immunopathol 43, 563–575 (2021). https://doi.org/10.1007/s00281-021-00867-8
2. Mandal A, Biswas N, Alam MN. Implications of xenobiotic-response element(s) and aryl hydrocarbon receptor in health and diseases. Hum Cell. 2023 Sep;36(5):1638-1655. doi: 10.1007/s13577-023-00931-5. Epub 2023 Jun 17. PMID: 37329424.
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