P(Lac)IQ RBS Aiia RBS LuxR ter ter luxpR RBS eyfp ter ter
We designed and built a kill switch mechanism that based on the activation of a lethal gene downstream of an inducible promoter that can be activated over time. In order to understand our system we started to test the first part of our genetic circuit , to test it we built the first part ( k1767001 ) driving expression of florescence protein . The here presented BioBrick consists of a constitutive promoter(I14032) driving expression of LuxR followed by a Lux promoter (R0062) , driving expression of YFP. The idea is to retrieve information about the expression of a fluorescent reporter downstream of the pLuxR promoter in absence and increasing concentrations of AHL. The scheme below explains how this BioBrick works :
P(Lac)IQ RBS Aiia RBS LuxR term term luxpR
parts registry and gBlocks ordered from idt
We designed and built a kill switch mechanism that is based on the activation of a lethal gene downstream of an inducible promoter that is activated over time. The presented part is the first module of our designed genetic circuit (BBa_K1767003). In order to characterize and understand each module of the genetic circuit we designed two composite parts that allow us to verify the (1) the degradation rate of AHL by AiiA and (2) induction of the pLux promoter by LuxR in presence of AHL with and without co-expression of AiiA. (1) The here presented BioBrick consists of a constitutive promoter driving expression of AiiA and LuxR followed by a Lux promoter which drives expression of a fluorescent reporter (here: yellow fluorescent protein). For a schematic overview see the figure below
medium RBS, YFP (-LVA), double terminator
Jay Blumling, Debra Lin, Madeleine Sheldon-dante, Fred Tan, MIT SMUG
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.
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.
enhanced yellow fluorescent protein derived from A. victoria GFP
Modificaitons to Clontech EYFP by Reshma Shetty
Transcriptional terminator consisting of a 64 bp stem-loop.
Conversion of the iGEM parts registry to SBOL2.1