Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jianyi Huang
Date created: 2016-10-12 11:00:00
Date modified: 2016-10-19 10:23:30
pTet+RBS+T7RNAP+Ter+pT7+RBS+tetX-GFP(fusion protein)+DT
| Types | DnaRegion |
| Roles | Reporter engineered_region |
| Sequences | BBa_K2150028_sequence (Version 1) |
Description
In this part, the first promoter pTet is constitutively on, producing T7 RNA polymerase(T7RNAP). T7RNAP can activate the T7 promoter, thus producing the fusion protein in large quantity. The 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.Notes
Two different terminators are used to avoid repeated sequence.Linker for fusion protein is carefully designed to avoid repeated sequence which may cause problems in assembly.
Source
T7RNAP:PCR amplified from Genome of BL21.pT7:PCR amplified from BBa_K525998.
As 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).