Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jianyi Huang
Date created: 2016-10-11 11:00:00
Date modified: 2016-10-17 08:48:10
pCon+RBS+tetR+DT+pTet+RBS+tetX-GFP(fusion protein)+DT
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K2150025_sequence (Version 1) |
Description
In this part, a fusion protein(referred to as tetX-GFP) of tetX(a tetracycline-degrading enzyme) and GFP combined with a 3*GGGGS linker is used to report the expression level of tetX. The first promoter(BBa_J23119) is constitutively on and can express tetR, a protein that can repress the downstream promoter pTet in the absence of tetracycline(Tc). As a result, no fusion protein tetX-GFP is expressed and no fluorescence can be detected. However, in the presence of Tc, tetR is repressed and repression over pTet is removed. This leads to the expression of tetX-GFP, degrading tetracycline and emitting fluorescence at the same time. This part can report activity of tetX in the presence of Tc by emission of green fluorescence.Notes
Linker for fusion protein is carefully designed to avoid repeated sequence which may cause problems in assembly.Source
pCon from BBa_J23119, RBS from BBa_B0034, tetR from BBa_C0040, DT from BBa_B0015,PTet from BBa_R0040As for fusion protein tetX-GFP, tetX is the same as (xxx), the protein linker is provided by our advisor Li Hua, and the GFP is from BBa_E0040. We combine this three parts using overlap extension polymerase chain reaction (OE-PCR).