Part:BBa_K5477024
pRAD27-AhR - receptor module
The pSTE12-AhR receptor module is designed as a composite with AhR BBa_K3793004 is expressed under the control of the pRAD27 promoter BBa_K5477002. AhR is a ligand-activated transcription factor that binds to a variety of environmental contaminants, including polycyclic aromatic hydrocarbons (PAHs), polychlorinated biphenyls (PCBs), and dioxins (1) (2). Upon binding these ligands, AhR translocates from the cytoplasm to the nucleus, where it forms a heterodimer with the Aryl hydrocarbon receptor nuclear translocator (ARNT). This complex then binds to xenobiotic response elements (XREs), initiating the transcription of detoxification genes, such as those encoding cytochrome P450 enzymes.
The pRAD27-AhR module was used for the biosensor device to detect environmental pollutants like PAHs and PCBs. Under the presence of these compounds, receptor modules including ARNT BBa_K5477025 and NCOA BBa_K5477026 will bind to XRE and initiate the transcription of NanoLuc in our reporter module BBa_K5477030. 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: BBa_K5477042
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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