TolC and Hemolysin B, C and D gene complex
This a composite part of BBa_K223055, BBa_K223056, BBa_K223057 that provides all necessary genes to create a channel protein in the extra cellular membrane of a prokaryotic cell. This channel can be used to export large organic molecules fused to a signal sequence, HlyAs
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.
GenBank Escherichia coli str. K-12 substr. MG1655 http://www.ncbi.nlm.nih.gov/nuccore/NC_000913.2/from=3176137&to=3177618&report=fasta Stanford iGEM Team 2009 altered the sequence restriction sites and unknown portions of the sequence.
TolC, along with HemolysinB, C, and D, for a channel in the cellular membrane to allow for the extracellular transport of HlyAs-fused protein sequences. The TolC receptor, in conjunction to the Hly genes, can serve as channel for transporting large proteins.
Front Flanking Sequence (HlyR, IS1, HlyC)
Compiled from several different papers: 1. Cloning of a cDNA Encoding an Aldehyde Dehydrogenase and Its Expression in Escherichia coli Xianshu Wang, Peter Penzes, and Joseph L. Napoli THE JOURNAL OF BIOLOGICAL CHEMISTRY. Vol. 271, No. 27, Issue of July 5, pp. 16288 ???16293, 1996 2. Escherichia coli CFT073 (GenBank) http://www.ncbi.nlm.nih.gov/nuccore/NC_004431.1/from=3418210&to=3418884&report=fasta
This front flanking sequence has three of the hemolysin parts - HlyR, Is1 and HlyC. In conjunction with rear flanking sequence, containing HlyB and HlyD, and TolC (part K223055), this part creates a channel protein to facilitate the extracellular export HlyAs-fused proteins.
Compiled from several different sources: 1. GenBank E. coli str. CFT073 http://www.ncbi.nlm.nih.gov/nuccore/NC_004431.1/from=3422041&to=3424164&report=fasta 2. GenBank E. coli str. CFT073 http://www.ncbi.nlm.nih.gov/nuccore/NC_004431.1/from=3424183&to=3425619&report=fasta
Along with parts BBa_K223055 and K223056, this part helps form the channel protein for the extracellular transport of HlyAs-fused protein.
Stanford iGEM Team 2009 altered sequences for internal restriction sites
Conversion of the iGEM parts registry to SBOL2.1