Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: paul james
Date created: 2013-09-19 11:00:00
Date modified: 2015-05-08 01:09:35
ompC w/GFP + (BglII+BamHI)restriction sites
| Types | DnaRegion |
| Roles | engineered_region Composite |
| Sequences | BBa_K1173502_sequence (Version 1) |
Description
ompF is a specific promoter which is only activated when phosphorylated OmpR (OmpR-P) is present. OmpR is phosphorylated by the protein Cph8 (form by two proteins, Cph1 and EnvZ), which is usually associated with osmotic regulation. Cph8 is also responsive to red light; exposure to red light halts the phosphorylation of OmpR. The lack of OmpR-P means the ompF promoter is not actiavted, so the genes which follow it are not transcribed. There is a GFP after ompF, which has the nucleotide sequence from BBa_E0040. The GFP will be produced in most generally used competant cells, as Cph8 is present in their outer membranes for osmotic regulation. To control the levels of GFP produced, we recommend the use of EnvZ deficient cells and introducing the genes which code for Cph8 on a LCN plasmid. This would allow control of GFP production using only red light.However, the novel concept of this part is that you may replace GFP with a gene of your choice. The GFP is flanked by restriction sites for BglII and BamHI. This allows you to "cut out" the GFP without interfering with the standard 3A assembly (EcoRI, XbaI, SpeI and PstI) and replace it with any gene, providing it also has the BglII and BamHI restriction sites. These can easily be added using traditional cloning using PCR primers. This will theoretically allow any gene to be controlled by the ompF promoter, and therefore exposure to red light.