Part:BBa_I746200:Experience

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User Reviews

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No review score entered. iliakats

This part is highly toxic to E. coli. According to the Cambridge 2007 wiki, who constructed this part, colonies only grew with this part driven by the weak promoter BBa_J23115, stronger promoters gave no colonies. In my own experience, E. coli transformed with Gibson assembly mix of this BioBrick driven by BBa_R0010 select for non-functional constructs where mutations were introduced by primer impurities. 9 positive clones were sequenced, out of those 8 habored deletions or point mutations introducing a stop codon at the beginning of the coding sequence, and 1 clone had a mutated RBS. A functional construct could only be obtained by making the weak BBa_J23114 promoter drive expression of this part. E. coli harboring this part driven by BBa_J23114 in a high-copy plasmid like pSB1C3 grow extremely slowly, normal growth only occurs when the functional construct is in a low-copy backbone like pSB3C5.

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