Upstream mutated chitoporin part fused with lacZ
The subparts were prepared: ChiP upstream part from Escherichia Coli W3110 genomic DNA by PCR and lacZ from part BBa_I732005 present in part registry.
ChiP encodes enterobacterial chitoporin required for uptake of chitin-derived oligosaccharides. This outer membrane porin is required for Salmonella and E.coli growth on chitooligosaccharides as a sole source of both carbon and nitrogen. It is only synthesized in the presence of these substrates. In nature it is regulated by sRNAs.In this biobrick we altered the binding site for ChiP regulator ??? ChiX. It was done in order to show that the regulatory system depends on the specific base pairing between ChiP and its regulator. The upstream part of this gene is fused with reporter gene lacZ to allow checking if the regulation works correctly.
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.
lacZ (encoding beta-galactosidase, full-length)
PCR amplified and standardized from BL21
LacZ encodes beta-galactosidase, an intracellular enzyme that cleaves the disaccharide lactose into glucose and galactose. Beta-galactosidase can also cleaves X-gal and ONPG into colorful products.
ChiP promoter and 5' UTR; RBS mutated AAGAGG-->AGGAGA
It was obtained by PCR from Escherichia coli W3110 genomic DNA.
ChiP encodes enterobacterial chitoporin required for uptake of chitin-derived oligosaccharides. This outer membrane porin is required for Salmonella and E.coli growth on chitooligosaccharides as a sole source of both carbon and nitrogen. It is only synthesized in the presence of these substrates. In nature it is regulated by sRNAa. We introduced the mutations in this sequence in the regulator binding site.
Conversion of the iGEM parts registry to SBOL2.1