Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Steven Chen
Date created: 2014-05-05 11:00:00
Date modified: 2015-05-08 01:14:05
FNR-dependent Hypoxia Sensor
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_M36667_sequence (Version 1) |
Description
This part utilizes fumarate nitrate reductase (FNR) transcription factor with E. Coli constitutive promoter, which regulates pepT7 promoter downstream with Gemini actuator. FNR and pepT7 promoter sequence are from Salmonella typhimurium, and the constitutive promoter, ribosome binding site, and terminator are generic E. Coli parts.FNR is regulated by O2: inactive when exposed to O2, and requires low O2 (hypoxia) and glutathione/cysteine desulfurase to reactivate, of which cysteine desulfurase is already present within E. Coli. Once activated, FNR induces the pepT7 promoter, which facilitates expression of the Gemini actuator. The lacZ portion of Gemini will produce beta-galactosidase, which can be qualitatively and quantitatively measured.
Notes
Modifications were made to the FNR sequence: GAG to GAA at position 418 and ACG to ACA at 421 to remove a restriction site, BbsI, but preserve the amino acid sequence.Source
1. Unden, Gottfried, and Jan Schirawski. "The Oxygen-responsive Transcriptional Regulator FNR of Escherichia Coli: The Search for Signals and Reactions."Molecular Microbiology 25.02 (1997): 205-10. Web.2. "Part:BBa_J23100.??? Registry of Standard Biological Parts. Web.
3. ???Part:BBa_B0015.??? Registry of Standard Biological Parts. Web.
4. Lombardo, Mary-Jane, Angela A. Lee, Tina M. Knox, and Charles G. Miller. "Regulation of the Salmonella Typhimurium PepT Gene by Cyclic AMP Receptor Protein (CRP) and FNR Acting at a Hybrid CRP-FNR Site."Journal Of Bacteriology 179.6 (1997): 1909-917. American Society for Microbiology. Web. 04 May 2014.
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J23100 const. promoter pRBS-SD1 FNR gene BBa_B0015 terminator pepT7 FNR-inducible promoter | 1,30 31,37 44,796 797,925 926,971 | feature/promoter promoter feature/rbs ribosome_entry_site feature/protein CDS stop_codon feature/stop promoter feature/promoter |