Part:BBa_K4769000:Design

Design Notes

pMiniMAD contains an internal BsaI cut site within the ampicillin resistance gene – this can be avoided when using BsaI by ensuring minimal homology of the internal 4 bp overhang with other designed overhangs and performing an additional final ligation step following Golden Gate assembly.

Alternatively, the BsaI recognition sequence could be removed by point mutation to a synonymous codon, by site-directed mutagenesis or using an additional internal set of primers.