pBAD/araC.luxABG.R0011N.luxCDE - bright
Part K1725350 is a reorganised luxABGCDE operon with pBAD promoter upstream luxA and K1725080)promoter upstream luxC generated by BioBrick assembly of pBAD.luxABG-bright (BBa_K1725350)and K1725080.luxCDE-bright (BBa_K1725351).
More information about the part.
Design of the RBS library for luxABG
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. We report that the final construct K1725352 (pBAD/araC.luxABG.K1725080.luxCDE) is the brightest that we found out of over 1 billion possible combinations.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 1205
Illegal NheI site found at 1780
Illegal NheI site found at 4057
- 21Illegal BglII site found at 6106
Illegal BamHI site found at 1144
Illegal XhoI site found at 1608
- 23COMPATIBLE WITH RFC
- 25Illegal AgeI site found at 979
- 1000Illegal BsaI site found at 3156
Illegal BsaI.rc site found at 5491
Illegal SapI.rc site found at 6659
Illegal SapI site found at 961