Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Eva D
Date created: 2013-09-22 11:00:00
Date modified: 2015-07-09 11:36:37
lacIq-T7-ccdB
| Types | DnaRegion |
| Roles | Coding CDS |
| Sequences | BBa_K1105000_sequence (Version 1) |
Description
This part contains a ccdB gene under control of a T7-promotor. CcdB is a bacterial toxin that interferes with topoisomerase unit gyrA. The toxin CcdB can be neutralized by its antitoxin CcdA. The T7-promoter allows for the controlled expression of CcdB.The goal of the UGent team 2013 is to improve the existing CIChE technique(Chemically inducible chromosomal evolution),developed by Tyo et al. in 2009. This was done by replacing the antibiotic resistance genes in the genome by CcdA antitoxin genes. This part was used to put pressure on the bacterial cells, in that way inducing chromosomal evolution.
Notes
We had problems cloning our part in the pSB1C3 backbone due to the high copy number of pSB1C3. In every attempt mutations occurred. This is probably caused by the high pressure leaky expression of the T7 promoter may put on the cells. In the case of leaky expression, mutants (no ccdB expression)have a huge growth advantage over cells containing the correct. This lead us to submit our part in the pSB6A1 backbone (low copy number).Source
This part (BBa_K1105000) comes from p10-LacIq-T7ccdB, a plasmid constructed by the lab Inbio-UGent. Originally, it comes from strains which are mini F-plasmid positive, such as E. coli F+.| Sequence Annotation | Location | Component / Role(s) |
| -35box -10box LacI LacIq pT7 LacO RBS-1 ccdB ccdB Start codon ccdB Stop codon | 44,49 67,72 108,230 231,1190 1526,1542 1547,1567 1600,1605 1613,1921 1613,1615 1919,1921 | feature/tag tag tag feature/tag CDS feature/cds CDS feature/cds feature/promoter promoter operator feature/operator feature/rbs ribosome_entry_site feature/cds CDS start_codon feature/start stop_codon feature/stop |