Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Isaac Bean
Date created: 2016-10-18 11:00:00
Date modified: 2016-10-19 05:11:50
RI7_FLAG_OXTR
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K2111000_sequence (Version 1) |
Description
This part is a chimeric receptor with a FLAG tag. It is composed of the ligand biding domain of the human oxytocin receptor, flanked by the N and C termini of the rat RI7 olfactory receptor. It was designed using the DIY receptor design guide created by the 2012 TU Delft iGEM team (http://2012.igem.org/Team:TU-Delft/receptordesign). This part is intended to express in the membrane of a yeast cell and - upon oxytocin ligand binding - activate the mating pathway and the inducible FUS1 promotor. This allows us to leverage a reporter protein under control of FUS1 to detect the presence of oxytocin.Notes
We added a FLAG tag to facilitate SDS-PAGE. We added the yeast kozak sequence tacaaa to improve translation and codon optimized for saccharomyce cerevisiae.Source
The human oxytocin receptor sequence was found in the NCBI database, codon optimized for saccharomyce cerevisiae using http://www.jcat.de/ (with additional options to avoid cleavage sites EcoR1, Pst1, Spe1, Xba1. The sequence for the rat RI7 OR was found on the 2012 TU Delft iGEM team page and in Rhadhika 2007 (http://www.nature.com/nchembio/journal/v3/n6/full/nchembio882.html).| Sequence Annotation | Location | Component / Role(s) |
| Kozak FLAG tag RI7 N-terminus Start Human Oxytocin Receptor Binding Site RI7 C-terminus Stop | 1,6 10,33 34,213 7,9 214,1017 1018,1116 1117,1122 | feature/rbs ribosome_entry_site feature/tag tag sequence_alteration feature/mutation start_codon feature/start non_covalent_binding_site feature/binding feature/mutation sequence_alteration feature/stop stop_codon |