Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: The Newcastle 2008 iGEM team
Date created: 2008-10-23 11:00:00
Date modified: 2015-06-09 09:00:20
Sensor for small peptide Subtilin
| Types | DnaRegion |
| Roles | Signalling engineered_region |
| Sequences | BBa_K104001_sequence (Version 1) |
Description
This part encodes a two-component system that can be used to sense the level of a small lantibiotic peptide called subtilin. It belongs to a series of parts that can be used to sense quorum sensing peptides from Gram positive organisms. In the Newcastle 2008 iGEM entry it was used a 'proof of concept' brick to test whether a peptide produced by one organism could be sensed by a different organism.The part contains two orfs arranged in an operon, SpaK and SpaR, that are driven by their native RBSs and the promoter PspaRK. Downstream of the SpaRK operon is terminator followed by a RBS and a promoter PspaS.
In response to subtilin, the sensor kinase SpaK will phosphorylate, and hence activate, the response regulator SpaR. The phosphorylated SpaR protein activates the PspaS promoter driving the expression of any downstream genes. In this brick there are no ORF's downstream of PspaS but the brick has been tested with gfp and mcherry by the Newcastle 2008 team.
Notes
A terminator was included after the spaRK to prevent read through. The native SpaRK promoter is controlled by the sigma factor sigma-H and induced at the end of exponential phase. The regulation by sigma-H was left unmodified.Source
The two component system, its promoter and RBS's, and the pSpaS promoter sequence are all derived from the genome of Bacillus subtilis ATCC 6633.| Sequence Annotation | Location | Component / Role(s) |
| Prom_spaRK RBS_spaR spaR RBS_spaK spaK ter_rrn0 Prom_spaS | 7,123 124,132 137,799 772,780 772,1953 1939,1957 2043,2120 | promoter feature/promoter feature/rbs ribosome_entry_site feature/cds CDS ribosome_entry_site feature/rbs feature/cds CDS stem_loop feature/stem_loop promoter feature/promoter |