Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Nandjafot MENDY
Date created: 2014-09-29 11:00:00
Date modified: 2015-05-08 01:10:18
P0 promoter-RBS B0032-sfGFP- Terminator B0015 - Pr promoter-DmpR
| Types | DnaRegion |
| Roles | Measurement engineered_region |
| Sequences | BBa_K1413001_sequence (Version 1) |
Description
This part consist on a sensor of phenolic compounds based on a transcription factorof the Ntrc family , DmpR.Found in Pseudomonas sp. strain CF 600, DmpR regulates expression from the Po promoter, which drives transcription from one single large operon for phenol degradation (dmpKLMNOPQBCDEFGHI).
With GFP attached to P0 promoter, it is then possible to evaluate presence of phenol by fluorescence analysis, if DmpR is expressed.
Thus this part is basically composed of [P0 promoter + sfGFP + Pr promoter + DmpR] and allows sensing of phenolic compounds thanks to a constitutively expressed DmpR that will activate expression of GFP.
We characterized this biosensor using phenol as effector.
Notes
We performed an assembly of the two parts by :-digesting BBa_K1031222 with EcorI and XbaI while digesting BBa_K1031211 by EcorI and SpeI.
-ligating the two digestion products.
BBa_K1031211 (Dmpr) had an extra Pst1 restriction site at position 226 (position 0 being EcoR1 restriction site).
We thus performed several trials of site directed mutagenesis and finally managed to modify a single nucletide in the Pst1 site, with respect to the amino acids sequence.
Source
This part is a composition of two parts from Peking iGEM 2013: BBa_K1031222 (sfGFP) & BBa_K1031211 (DmpR)Professor V.Shringler provided 2013 Peking iGEM Team with pVI401 composed of Pr-DmpR on vector pVI39.
| Sequence Annotation | Location | Component / Role(s) |
| P0 Promoter RBS Junction RBS B0032 RBS Junction sfGFP Terminator B0015 SpeI/XbaI mixed site PR Promoter DmpR BBa_K1413001 | 1,171 172,179 180,192 193,198 199,929 930,1058 1059,1066 1082,1155 1156,2847 1,2847 | promoter feature/promoter feature/misc sequence_feature feature/rbs ribosome_entry_site sequence_feature feature/misc feature/cds CDS sequence_feature feature/misc sequence_feature feature/misc feature/promoter promoter CDS feature/cds feature/BioBrick engineered_region |