This part includes the S.aureus Agr quorum sensing sensor which recognizes the oligopeptide. The FepA pore allows for the oligopeptide to diffuse through the outer-membrane.
RBS (Elowitz 1999) -- defines RBS efficiency
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix. 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.
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.
AIP sensor infrastructure (agrC + agrA from S. aureus with RBSes)
The original sequences for agrC and agrA were taken from the ''S. aureus'' Sequencing Project (results for strain NCTC8325) at Oklahoma University (http://www.genome.ou.edu/staph.html). The sequences were then codon-optimised for ''E. coli'' using GeneDesigner from DNA2.0 (http://www.dna20.com), and then synthesised by that company.
This part was made in order to transfer the S. aureus oligopeptide-based quorum sensing system into a BioBrick-compatible signalling mechanism. In the natural system, the signalling oligopeptide (termed AIP) is made from AgrD by the membrane-located enzyme AgrB. It is then detected by the membrane-located AgrC, which phosphorylates AgrA which then has DNA-binding activity and upregulates transcription of the promoters termed P2 and P3 in the agr locus. There are four known variants of AIP with different molecular structures and cross-inhibitory activity; this BioBrick generates group I AIP. This part contains the coding regions for group I agrC and agrA (in that order) from ''S. aureus'' strain NCTC8325, prefixed by the B0034 RBS and separated by a BamHI site.
FepA L8T Mutant - Large Diffusion pore for E. coli outer membrane.
The mutants were developed by Dr Salete Newton working in Professor P. Klebba's lab at the university of Oklahoma. They were kindly sent to us by Professor Klebba.
FepA is an outer membrane receptor protein for Escherichia coli involved in the uptake of the ferric enterobactin siderophore. This part is a deletion mutant of the wild type FepA, which has been shown to convert the protein into an open diffusion channel, similar to a porin. It allows larger molecules through than normally would be permitted.
The original mutants had an EcoR1 restriction site within their coding sequence. This had to be removed via site directed mutagenesis, using a megaprimer based method.
Conversion of the iGEM parts registry to SBOL2.1