Constitutive expression of GFP-REACh1 fusion protein
The part is combined from other BioBricks
This part expresses a E0040.K1319001 fusion protein (EGFP-REACh1) behind a J23101 promotor.
We designed this part to be very similar to I20260 and the other fusion parts (K1319014 and K1319015).
Florian Gohr, Arne Zimmermann, Michael Osthege
RBS.3 (medium) -- derivative of BBa_0030
Weak1 RBS based on Ron Weiss thesis. Strength is considered relative to <bb_part>BBa_B0030</bb_part>, <bb_part>BBa_B0031</bb_part>, <bb_part>BBa_B0033</bb_part>.
Varies from -6 to +1 region from original sequence to accomodate BioBricks suffix ("RBS-2" in figure 4-14 of thesis). Contact info for this part: Brian Chow
Vinay S Mahajan, Voichita D. Marinescu, Brian Chow, Alexander D Wissner-Gross and Peter Carr IAP, 2003.
This is the sequence of the SpeI/XbaI scar for RBS-CDS junctions in BioBricks standard assembly.
This is a shorter scar to ensure proper spacing between the RBS and CDS.
XbaI/SpeI mixed site. Simply used to aid in entry of parts into the registry.
This is the scar produced by assembly using RFC 25. If you are assembling a composite part using RFC 25, you can insert this part and specify blunt assembly to get the desired sequence.
TEV cleaves the following AA sequence with high specificity: * Glu-Asn-Leu-Tyr-Phe-Gln↓Gly but: * Glu-Asn-Leu-Tyr-Phe-Gln↓Ser is also reported Note, the part suggested by the Voigt lab, also contains 2 additional 1xGly flanks for increased accessibility. See also: * BBa_I712077 (TEV N-term) * BBa_I712078 (TEV C-term) * BBa_I712016 (myri+TEV site, Slovenia team igem2007) * BBa_J64007 (Dan Widmaier, Voigt lab) References =========== Dougherty et al. (1989): Molecular genetic analysis of a plant virus polyprotein cleavage site: a model. PMID: 2669323
* gene synthesis * codon optimized for E. coli
RFC[25]-compatible dark quencher based on K1319000 (E0030)
The part was created by site-directed mutagenesis on K1319000 that we previously created from E0030
This part is a RFC[25] dark quencher that is based upon K1319000 (the RFC[25] version of E0030/EYFP). Two mutations were introduced that eliminated fluorescence: * L90I * Y145W =References= * Ganesan S, Ameer-Beg SM, Ng TT, Vojnovic B, Wouters FS. A dark yellow fluorescent protein (YFP)-based Resonance Energy-Accepting Chromoprotein (REACh) for F??rster resonance energy transfer with GFP. Proc Natl Acad Sci U S A. 2006 Mar 14;103(11):4089-94. Epub 2006 Mar 6. PubMed PMID: 16537489; PubMed Central PMCID: PMC1449651. [http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1449651/?report=classic PubMed Central]
The Y145W mutation was introduced to eliminate fluorescence of the protein.
Double terminator consisting of BBa_B0010 and BBa_B0012
This sequence is produced by an assembly method.
This sequence is produced by RFC10-assembly of a RFC25-CDS with a RFC10-prefixed part.
The resulting sequence is a combination of 9 nucleotides from the RFC25 prefix (acc ggt taa) and the RFC10 scar (tactagag).
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