"IPTG bomb" kill the E.coli by coding Ccdb toxin protein. The Ccdb come from BBa_K581008(RBS+ccdB+Termintor),and T5-lac Promoter from BBa_K592008
A system designed to kill E.coli when IPTG added . IPTG can efficiently trigger a large numumber of Ccdb protein .And Ccdb protein a can combine with E.coli ，So it can effectively kill the E.coli.
We have been thinking of a way, that way we could eliminate E.coli. Now we Ccdb protein-bound T5-lac and make the controls to suit our needs.
T5-lac promoter is a hybrid promoter made from the phage T5 early promoter and lac-operon. It contains three LacI binding sites and remains repressed in LacIq strains where LacI is expressed in high levels. It is inducible by IPTG, and recognizable by E.coli RNA polymerase. Compared to the T7 promoters, T5-lac promoter doesn't require the co-expression of phage polymerase.
The most common DH5alpha and TOP10 used in assembly are not LacI-constitutive (LacIq) strains. Thus, any ORF placed downstream to this promoter will be massively transcribed. Therefore, it's recommended to employ low-copy number plasmids or even LacIq strains when adding genes to this promoter.
This is the cell death protein coding sequence under the control of a relatively weak RBS. It is replicable in non-ccdB resistant strains.
Use a relatively weak RBS so the coding sequence would be replicable in non-ccdB resistant strains under the putative promoter in VF region.
Double terminator consisting of BBa_B0010 and BBa_B0012
RBS.4 (weaker) -- derivative of BBa_0030
Weaker RBS based on Ron Weiss thesis. Strengths relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0032</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-3" in figure 4-14 of thesis). No secondary structures are formed in the given RBS region. Users should check for secondary structures induced in the RBS by upstream and downstream elements in the +50 to -50 region, as such structures will greatly affect the strength of the RBS. Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
Coding region for the ccdB (control of cell death) gene.
Derived from the TE terminator of T7 bacteriophage between Genes 1.3 and 1.4 <genbank>V01146</genbank>.
Transcription terminator for the E.coli RNA polymerase.
Suggested by Sri Kosuri and Drew Endy as a high efficiency terminator. The 5' end cutoff was placed immediately after the TAA stop codon and the 3' end cutoff was placed just prior to the RBS of Gene 1.4 (before AAGGAG). Use anywhere transcription should be stopped when the gene of interest is upstream of this terminator.
Transcriptional terminator consisting of a 64 bp stem-loop.
Conversion of the iGEM parts registry to SBOL2.1