Team:Glasgow/Composite Part

pBAD/araC.luxABG - bright

Part K1725350 is a luxABG gene assembly under the pBAD promoter generated by BioBrick assembly of luxA (BBa_K1725200), luxB (BBa_K1725201) and luxG (BBa_K1725205) ribosome binding site (RBS) libraries. RBS BBa_K1725305, BBa_K1725309 and BBa_K1725302 are upstream of luxA, luxB and luxG genes, respectively. The assembly has been shown to induce high levels of bioluminescence in E. coli in a presence of decanal.

For the construction of the RBS library, a master sequence based on the RBS B0032 was used. 4 nucleotides within the actual ribosome binding site were randomised giving 32 different B0032-derived RBS variants. The predicted efficiency of each RBS library member was estimated using RBS Library Calculator. Originally, RBS Library was designed for every gene in luxABGCDE operon (BBa_K1725352) in order to optimise bioluminescence in E. coli. 32 different BioBrick RBS variants were incorporated upstream of each of the lux genes by PCR, and different libraries were then combined together by BioBrick assembly. This method could potentially produce over 1 billion different variants of the lux operon. Just for the luxABG assembly, there are over 32000 different variants. K1725350 is the brightest colony from the >32000 variants that we found. See our RBS library page for more information.


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