Part:BBa_K4675005

This plasmid is a further development of the iGEM Münster 2022's level 2 shuttle vector pSB3CY_aeBlue (BBa_K4188001) for Sacchaomyces cerevisiae. We adjusted it, so it can be used as a shuttle vector for Komagatalella phaffii (Pichia pastoris). We exchanged the existing 2μ Ori with the origon of replication PARS for K. phaffii. The aeBlue chromoprotein (BBa_K864401) flanked by the promoter (BBa_J23110) and the terminator (BBa_B0015) can be replaced with a selection marker using the restriction enzyme BsmBI. The colonies will show a pink color if the aeBlue Chromoprotein is replaced, otherwise they apper in a dark blue color. If the mRFP is exchanged with the expression casette, the colonies appear in a white color. This changing color system makes it easy to check for plasmids with inserted selection markers and expression casettes. If the selection marker needs to be changed later on, the restriction enzymes NotI, SpeI and BamHI can cut out the marker.

As a ready to use vector, we inserted the selection marker for the antibiotic Zeocin. pSB3CYP_Zeo (BBa_K4675008)