Part:BBa_K5477025
pRET2 - ARNT receptor module
The pRET2-ARNT receptor module is a composite part designed to control the expression of ARNT (Aryl hydrocarbon receptor nuclear translocator) | BBa_K3793005 under the regulation of the pRET2 promoter BBa_K5477000. ARNT forms heterodimerization with the Aryl hydrocarbon receptor (AhR) in the nucleus. This heterodimer regulate gene transcription in response to xenobiotics (1) (2). This complex then binds to xenobiotic response elements (XREs) in the promoter regions of detoxification genes (1) (2) (3) (4).
This module supports the AhR-mediated response to toxic ligands like PAHs, dioxins and PCBs by binding to the XRE in the reporter module facilitating the transcription of NanoLuc BBa_K5477030. The composite part was cloned using the method of USER-cloning into YCp-H. YCp-H is a centromeric plasmid used in yeast that includes a HIS3 marker, allowing for selection in histidine auxotrophic yeast strains. Like other CEN plasmids, YCp-H contains a CEN sequence, ensuring that the plasmid replicates and segregates similarly to yeast chromosomes. This results in a low copy number (typically one to two copies per cell), providing stable maintenance of the plasmid. This composite part was used in the following devices: BBa_K5477041 and BBa_K5477042
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916
Illegal SpeI site found at 1760 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916
Illegal NheI site found at 182
Illegal SpeI site found at 1760 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916
Illegal SpeI site found at 1760 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 810
Illegal EcoRI site found at 1916
Illegal SpeI site found at 1760
Illegal AgeI site found at 37 - 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. Goedtke L, Sprenger H, Hofmann U, Schmidt FF, Hammer HS, Zanger UM, Poetz O, Seidel A, Braeuning A, Hessel-Pras S. Polycyclic Aromatic Hydrocarbons Activate the Aryl Hydrocarbon Receptor and the Constitutive Androstane Receptor to Regulate Xenobiotic Metabolism in Human Liver Cells. Int J Mol Sci. 2020 Dec 31;22(1):372. doi: 10.3390/ijms22010372. PMID: 33396476; PMCID: PMC7796163.
3. Kafafi SA, Afeefy HY, Ali AH, Said HK, Kafafi AG. Binding of polychlorinated biphenyls to the aryl hydrocarbon receptor. Environ Health Perspect. 1993 Oct;101(5):422-8. doi: 10.1289/ehp.93101422. PMID: 8119253; PMCID: PMC1519849.
4. Huang G, Elferink CJ. A novel nonconsensus xenobiotic response element capable of mediating aryl hydrocarbon receptor-dependent gene expression. Mol Pharmacol. 2012 Mar;81(3):338–47.
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