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.
Double terminator consisting of BBa_B0010 and BBa_B0012
Medium RBS based on Ron Weiss thesis. Strength considered relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0032</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-1" 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 Cho
prtD_prtE_prtF from erwinia chrysanthemi
Gene Bank ID: Erwinia chrysanthemi M60395.1
This is the 3 genes: prtD, prtE, prtF, that are from a type I Erwinia chrysanthemi secretion system.
Synthesized to be bio-bricked, keeps original protein sequence and ribosome binding site.
Secretion Signal and Streptavidin Binding Tags
Nanotag (bases 1-48): From Lamla and Erdmann (http://www.ncbi.nlm.nih.gov/pubmed/14680960). Secretion tag (last 181 AA): Gene Bank ID Erwinia chrysanthemi M60395.1
This part is the vector into which afp is inserted to tag it for purification using the IPP system. It contains an NheI restriction site that allows for the insertion of the desired protein coding sequence (NheI is compatible with XbaI and SpeI). This Target Vector part contains two tags: a secretion tag that causes the protein to be exported to the extracellular space, and a nanotag that causes the protein to bind to stretavidin (or some analog). For this part to function properly, it must be combined with a promoter and RBS, and used in E.coli strain UW5alpha, which has two other plasmids containing the other genes required for proper secretion and display of the protein to be purified on the outer membrane.
NheI was chosen as the protein insertion site because it is compatible with digested XbaI and SpeI restriction sites. The TEV sites and HIS tags flanking the NheI site are present to separate the inserted protein from the added tags. GSA linkers were used to separate the various features of the part.
1) The cumate gene-switch: a system for regulated expression in mammalian cells Alaka Mullick, Yan Xu, Ren?? Warren, Maria Koutroumanis, Claire Guilbault, Sophie Broussau, F??lix Malenfant, Lucie Bourget, Linda Lamoureux, Rita Lo, Antoine W Caron, Amelie Pilotte and Bernard Massie ( BMC Biotechnology 2006, 6:43 doi:10.1186/1472-6750-6-43 ) 2. Novel, Versatile, and Tightly Regulated Expression System for Escherichia coli Strains Young J. Choi, Lyne Morel, Teffanie Le Fran??ois Denis Bourque, Lucie Bourget, Denis Groleau, Bernard Massie and Carlos B. M??guez (Applied and Environmental Microbiology, Aug. 2010, p. 5058???5066 Vol.76 doi:10.1128/AEM.00413-10)
￼￼￼This promoter is composed by phage T5 promoter and Cumate operator. It is repressed in the presence of CymR protein which binds the Cumate Operator.
We added the standard iGEM prefix and suffix in order to clone it into the standard iGEM vector pSB1C3
T5LacO-LPP-OmpA-scFv (scFv 54.6 antinorovirus)-6HIS-TT
DNA synthesis. 1.???Transport and anchoring of 8-lactamase to the external surface of Escherichia coli??? J. A. Francisco, C. F. Earhart and G. Georgiou. Proc. Natl. Acad. Sci. USA Vol. 89, pp. 2713-2717, April 1992 Biochemistry. 2. ???Recombinant norovirus-specific scFv inhibit virus-like particle binding to cellular ligands??? K. Ettayebi and M. E. Hardy. Published: 31 January 2008 in Virology Journal 2008, 5:21?
This construct is designed for the expression of an already descibed engeneered antinorovirus (NoV) monoclonal antibody (mAb 54.6) in fusion with LPP-OmpA. The antibody is expressed in a single chain fragment variable (scFv) format containing light (VL) and heavy (VH) variable domains separeted by a flexible peptide linker. It has already been reported that the scFv 54.6 binds a native recombinant NoV particles (VLPs) and inhibits VLP interaction with cells. LPP-OmpA functions as a leader sequence and an anchor to display the scFv ot the bacterial surface.The construct consistes of T5 Lac Operator (Bba_K875002), ribosomal binding site, LPP-OmpA-scFv 54.6 antinorovirus, Histidine tag (6HIS), Terminator (B0015).
This coding sequence was optimized for the expression in prokaryotic cells. It has a Histidine tag.
The sequence was obtained by applying all mutations that upregulated AraC binding and subsequent promoter activity listed in reference [1].
pBAD is an E.coli promoter that is induced by L-arabinose. K206000 is a mutagenized pBAD promoter that is responsive to lower concentrations of arabinose than wild type (<partinfo>I13453</partinfo>) and, additionally, exhibits a higher maximum of protein expression as measured by coupling to a fluorescent reporter.
There were no special design considerations.
A toxic protein originating from pseudomonas aeruginosa that has been shown to arrest growth in both prokaryotic and eukaryotic cells when expressed intracelluarly. It is a substrate p. aruginosas type 6 secretion system
mention something about promoter needs to have no leaky expression for succesful cloning
Matthew Coyne, Ingrid Swanson, Jesa Landis, Matthew Harger
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.
This is one of the 3 genes: prtD, prtE, prtF, that are from a type I Erwinia chrysanthemi secretion system.
Synthesized to be bio-bricked, keeps original protein sequence.
Synthesized to be bio-bricked, keeps original protein sequence and rhibosome binding site.
Conversion of the iGEM parts registry to SBOL2.1