Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Sofia Chinea
Date created: 2016-09-12 11:00:00
Date modified: 2016-10-20 11:47:25
CpxR binding site attached to a yellow-green color protein (YGCP) acts as a neutral pH indicator.
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_K2097000_sequence (Version 1) |
Description
CpxA-CpxR is a two-component mechanism that is activated at pH 7.4 and repressed at pH 6.0. CpxA is an intermembrane protein that autophosphorylates at a certain external pH, CpxR (a kinase) then gets phosphorylated by CpxA and acts as a transcription factor. This system originally is a transcription factor for the virF gene, but we replaced virF with the Reporter. This reporter is a YGCP (Bba_K1033916). The original sequence was found in Shigella sonnei, but E. coli has a homolog of these proteins so all that is required on the construct is the appropriate prefix/suffix and CpxR binding site.Notes
The sequence directly from the papers - "Involvement of cpxA, a Sensor of a Two-Component Regulatory System, in the pH-Dependent Regulation of Expression of Shigella sonnei virF Gene" (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC177285/pdf/1775062.pdf) and "Identification of cpxR as a Positive Regulator Essential for Expression of the Shigella sonnei virF Gene" (http://www.ncbi.nlm.nih.gov/pmc/articles/PMC107317/pdf/jb003522.pdf) - was not used. We used the loci detailed in the papers and found the CpxR binding sequence in the GenBank to construct the sequenced used in the experiments.Source
The two parts together (CpxR binding site - YGCP) were ordered from IDT.The original sources for the CpxR binding site came from:
"Involvement of cpxA, a Sensor of a Two-Component Regulatory System, in the pH-Dependent Regulation of Expression of Shigella sonnei virF Gene"
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC177285/pdf/1775062.pdf
"Identification of cpxR as a Positive Regulator Essential for Expression of the Shigella sonnei virF Gene"
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC107317/pdf/jb003522.pdf
GenBank used: http://www.ncbi.nlm.nih.gov/nuccore/CP000039.1?&feature=CDS
The YGCP (Bba_K1033916) comes from the iGEM Registry.
| Sequence Annotation | Location | Component / Role(s) |
| Cpx-R Binding Site RBS Bba_K1033916 | 1,82 83,95 101,793 | feature/binding non_covalent_binding_site feature/rbs ribosome_entry_site feature/BioBrick engineered_region |