BBa_K314204
BBa_K314204 Version 1 Component
This part has been discontinued.
Please consider using BBa_K314300 as a replacement.
Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Laura Austin
Date created: 2010-10-21 11:00:00
Date modified: 2015-05-08 01:11:55
p4T6S::T7
| Types | DnaRegion |
| Roles | Plasmid plasmid |
| Sequences | BBa_K314204_sequence (Version 1) |
Description
A fosmid (F' BAC) which encodes the HSI-I Type VI Secretion locus of Pseudomonas aeruginosa. The locus has been engineered using homologous recombination to replace native promoters with robust T7 promoters. The two promoters are contained in the same intergenic region, with one on the positive strand and one on the negative.We are using this plasmid to express a Type VI Secretion System in the BL21(DE3) strain of E. coli which has an IPTG inducible T7 RNA polymerase. The purpose of the system is to create a probiotic system, creating a secretion system for a toxin/antitoxin system (BBa_K314200-BBa_K3142003).
The system allows for protein delivery directly from the donor cell into the periplasm or cytosol of the recepient organism. While we are using this system to deliver a toxin, other protein effectors might be used for a range of applications.
Notes
The size of the gene locus was too large to effectively BioBrick; there are several NotI and other restriction sites in crucial parts of necessary genes.Source
Original source was from P. aeruginosa PA01 genome library--fosmid O24--from Mike Jacobs at University of Washington Department of Immunology.| Sequence Annotation | Location | Component / Role(s) |
| ppkA pppA icmF1 fha1 RBS T7 promoter T7 promoter RBS hcp1 clpV1 VgrG1 | 13050,16148 16156,16884 17571,20876 24313,25806 25812,25817 25825,25844 25876,25894 25900,25905 29141,29629 34011,36719 36766,38697 | feature/protein CDS feature/protein CDS CDS feature/protein CDS feature/protein ribosome_entry_site feature/rbs feature/promoter promoter feature/promoter promoter feature/rbs ribosome_entry_site CDS feature/protein CDS feature/protein CDS feature/protein |