Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Jim Rose
Date created: 2011-07-04 11:00:00
Date modified: 2015-05-08 01:12:59
Fast-Fusion GFP-LacZa Reporter (RecA cleavage with Medium linker)
| Types | DnaRegion |
| Roles | Composite engineered_region |
| Sequences | BBa_K648024_sequence (Version 1) |
Description
This is a fast-acting reporter using the LacZa gene (BBa_K648012) fused to GFP (BBa_K648013). In its normal un-cleaved state the polymerization ability necessary for enzymatic action is inactivated. Once the fusion protein is cleaved by the RecA protein, which is activated by the presence of damaged DNA, the B-gal enzyme encoded by the LacZa gene is able to convert X-gal to galactose and 5-bromo-4-chloro-3-hydroxyindole, the latter of which is oxidized to the dark blue product 5,5'-dibromo-4,4'-dichloro-indigo. This reporter is able to produce a visible blue color response in a matter of minutes.This version uses the RecA cleavage site (BBa_K648009) followed by a 6 amino acid flexible fusion protein linker (BBa_K648005). The linker is attached to the N-terminus of the LacZa gene.
Notes
All parts were assembled via standard 25 assembly methods.Source
The design of this part was inspired by the Imperial College London 2010 iGEM Team's Fast-Response module, part BBa_K316007.| Access | Instance | Definition |
| public public public public public public public | BBa_J23100 BBa_B0034 BBa_K648013 BBa_K648009 BBa_K648007 BBa_K648012 BBa_B0015 | BBa_J23100 BBa_B0034 BBa_K648013 BBa_K648009 BBa_K648007 BBa_K648012 BBa_B0015 |
| Sequence Annotation | Location | Component / Role(s) |
| BBa_J23100 BBa_B0034 BBa_K648013 BBa_K648009 BBa_K648007 BBa_K648012 BBa_B0015 | 1,35 44,55 64,798 807,827 836,853 862,1089 1098,1226 | BBa_J23100 BBa_B0034 BBa_K648013 BBa_K648009 BBa_K648007 BBa_K648012 BBa_B0015 |