BBa_K2111000

BBa_K2111000 Version 1

Component

Source:
http://parts.igem.org/Part:BBa_K2111000
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
igem#status
Planning
 
synbiohub#ownedBy
user/james
 
synbiohub#ownedBy
user/myers
 
synbiohub#topLevel
BBa_K2111000/1