the research leading to these results has received funding from the innovative medicines initiative joint undertaking project chem21 under grant agreement no . 11  5360 , resources of which are composed of financial contribution from the european union 's seventh framework programme ( fp7/2007 - 2013 ) and efpia companies in kind contribution . we gratefully acknowledge the austrian science fund ( fwf ) project number w901 ( dk  molecular enzymology graz ) for funding and support from nawi graz .
nuclear localization sequences ( nlss ) are required for the import of proteins in the nucleus of eukaryotes . 
 however many proteins from bacteria or bacteriophages are used for basic studies in molecular biology , to generate synthetic genetic circuits or for genome editing applications . 
 prokaryotic recombinases , crispr - associated proteins such as cas9 or bacterial and viral polymerases require efficient nlss to function in eukaryotes . 
 the yeast pichia pastoris is a widely used expression platform for heterologous protein production , but molecular tools such as nlss are limited . 
 here we have characterized a set of 10 nlss for p. pastoris , including the first endogenous nlss ( derived from p. pastoris proteins ) and commonly used heterologous nlss . 
 the nlss were evaluated by fusing them in n- and c - terminal position to an enhanced green fluorescent protein showing pronounced differences in fluorescence levels and nuclear targeting . 
 thereby we provide a set of different nlss that can be applied to optimize the nuclear import of heterologous proteins in p. pastoris , paving the way for the establishment of intricate synthetic biology applications .