Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Bianca Kapoor, Sally Winkler
Date created: 2011-12-10 12:00:00
Date modified: 2015-05-08 01:14:04
Cadmium Sensor
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_M36246_sequence (Version 1) |
Description
This part was designed to promote the CadC gene, taken from E.Coli, constitutively with a bicistronic ribosome binding site. CadC serves as a transcription factor for the PCad promoter sequence. PCad serves to produce a PoPs signal that can then be used as an input for a sensor device.In order to detect Cadmium, we used a cadmium sensor from E.Coli. The genome of E.Coli codes for three genes, CadC, CadA, and CadB which allow for some resistance to cadmium. CadC, BBa_M36244, is a transcription factor that along with cadmium binds to a promoter, called Pcad, BBa_M36245, that promotes CadA and CadB. By using Pcad as a promoter, we can control the expression of any gene that follows it, using cadmium. To express Pcad, we also need CadC. We constitutively expressed CadC with a medium strength promoter from biofab.org, apFAB64. Following the promoter is a bicistronic ribosome binding site, BD2, BBa_M36282. CadC is already produced in E.Coli in small amounts, and more so with cadmium present. Our plasmid???s Pcad promoter may use either the CadC expressed from our plasmid or expressed naturally in E.Coli. Either way, Pcad will sense the presence of cadmium. After CadC, we placed a strong terminator, apFAB381.
Notes
During design, we used a medium strength promoter to ensure the promoter did not over work the cell.Source
Watson N., et al, (1992) Identification of Elements Involve in Transcriptional Regulation of the Escherichia coli cad Operon by External pH, Journal of Bacteriology, Vol 174, p. 530-540.biofab.org
Keselr, I., Collado-Vides, J., Santos-Zavaleta, A., Peralta-Gil, M., Gama-Castro, S., Muniz-Rascado, L., Bonavides-Martinez, C., Paley, S., Krummenacker, M., Altman, T., Kaipa, P., Spaulding, A., Pacheco, J., Latendresse, M., Fulcher, C., Sarker, M., Shearer, A., Mackie A., Paulsen, I., Gunsalus., and Karp, P., (2010), EcoCyc: a comprehensive database of Escherichia coli biology.
Escherichia coli str. K-12 substr. MG1655 chromosome, complete genome. NCBI Reference Sequence: NC_000913.2.
| Access | Instance | Definition |
| public public public public public | BBa_M36247 BBa_M36282 BBa_M36244 BBa_M36248 BBa_M36245 | BBa_M36247 BBa_M36282 BBa_M36244 BBa_M36248 BBa_M36245 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_M36247 BBa_M36282 BBa_M36244 BBa_M36248 BBa_M36245 | 1,37 46,133 140,1678 1687,1768 1777,1911 | BBa_M36247 BBa_M36282 BBa_M36244 BBa_M36248 BBa_M36245 |