As for the pGlowroplast I'm having partly synthesized at the moment (yes, they are slowly :D ):
I noticed that the genomic Vibrio fischeri fragment in pVIB (containing the Lux operon) is NOT from Vibrio fischeri ES114 but from starain MJ11. Had to learn it the hard way, restriction with BglII and SalI did not yield
BglII---RBS-LuxC-RBS-LuxD-RBS-LuxA-RBS-LuxB-RBS-LuxE-RBS-LuxG-terminator-salI
(roughly 6,5 kbp)
But rather
BglII--part of LuxD-RBS-LuxA-RBS-LuxB-RBS-LuxE-RBS-LuxG-terminator-salI
(~ between 4 and 5 kbp)
Fortunately recognized it on a gel before taking it as the fragment.
-- I noticed that the genomic Vibrio fischeri fragment in pVIB (containing the Lux operon) is NOT from Vibrio fischeri ES114 but from starain MJ11. Had to learn it the hard way, restriction with BglII and SalI did not yield
BglII---RBS-LuxC-RBS-LuxD-RBS-LuxA-RBS-LuxB-RBS-LuxE-RBS-LuxG-terminator-salI
(roughly 6,5 kbp)
But rather
BglII--part of LuxD-RBS-LuxA-RBS-LuxB-RBS-LuxE-RBS-LuxG-terminator-salI
(~ between 4 and 5 kbp)
Fortunately recognized it on a gel before taking it as the fragment.
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