Part:BBa_M50497:Design
uvrY Biosensor with lux Sp1 and luxSp2 Promoters
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 129
Illegal EcoRI site found at 594 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 129
Illegal EcoRI site found at 594 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 129
Illegal EcoRI site found at 594
Illegal BamHI site found at 803 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 129
Illegal EcoRI site found at 594 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 129
Illegal EcoRI site found at 594 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The two promoters needed to be inserted upstream of GFP so that upon activation the fluorescent protein would be produced accordingly.
Source
The two inducible luxS promoters come from Udekwu (2010), isolated from Escherichia coli. The GFP reporter comes from the iGEM repository.
References
https://parts.igem.org/Part:BBa_M50494
https://parts.igem.org/Part:BBa_M50491
Arindam M., et al. (2016). Integration of AI-2 Based Cell-Cell Signaling with Metabolic Cues in Escherichia coli. PLoS ONE 11(6): e0157532. https://doi.org/10.1371/journal.pone.0157532.
Federle, M.J. (2009). Autoinducer-2-Based Chemical Communication in Bacteria: Complexities of Interspecies Signaling. Contrib Microbiology 16: 18-32. Doi: 10.1159/000219371.
Pernestig, A.K., Melefars, O., and Georgellis, D. (2000). Identification of UvrY as the Cognate Response Regulator for the BarA Sensor Kinase in Escherichia coli. Journal of Biological Chemistry 276: 225-231. www.jbc.org/content/276/1/225.long.
RegulonDB. luxS operon and associated TUs in Escherichia coli K-12 genome. http://regulondb.ccg.unam.mx/operon?term=ECK120023629&format=jsp&organism=ECK12. Online.
Zere, T.R., et al. (2015). Genomic Targets and Features of BarA-UvrY (-SirA) Signal Transduction Systems. PLoS ONE 10(12): e0145035. Doi:10.1371/journal.pone.0145035.