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Revision as of 15:15, 31 October 2017

We will first create a yeast-E. Coli shuttle vector by putting ArgArs and PARS2 genes into psB1C3mut. Then, to create our centromeric plasmid, we must identify the minimal, essential region of the Pichia centromere that, when incorporated into a plasmid backbone, will allow the plasmid to mimic a Pichia chromosome during replication. This region can then be incorporated into Pichia backbones to convert them into centromere plasmids, which transform with high efficiency, have low and constant copy numbers, and are stably maintained without integration. Ultimately, we will contribute an important foundational tool for flexible, creative, and impactful cloning in yeast. If successful, the centromeric plasmids will serve a multitude of purposes in the bioengineering and industrial fields.