Table 1.

DNA parts used in expression cassette construction.

DNA Part	Characteristics	References
Promoters
pTEF	constitutive; native	Muller et al., 1998
pTDH1, pGPM1, pFBAIN	constitutive; native	Hong et al., 2012
pEXP1, pGPAT, pGPD	constitutive; native	Xue et al., 2013
pGAP, pACL2	inducers were not determined; native	Wong et al., 2017
pXPR2	inducible by peptones; native	Ogrydziak and Scharf, 1982
pPOX2	inducible by fatty acids and alkanes; repressed by glucose and glycerol; native	Juretzek et al., 2000
pPOT1	inducible by fatty acids and alkanes; repressed by glucose and glycerol; native	Juretzek et al., 2000
pLIP2	inducible by fatty acids and alkanes; native	Sassi et al., 2016
pICL	inducible by ethanol, fatty acids, and alkanes; native	Juretzek et al., 2000
pYAT1	induced by nitrogen-limited conditions; native	Xue and Zhu, 2012
hp4d	hybrid promoter derived from pXPR2; contains four copies of UAS1XPR2 fused upstream from a minimal core LEU2 promoter; growth phase dependent; hybrid	Madzak et al., 2000
n UAS1XPR2-LEU	hybrid promoter derived from pXPR2; core minimal LEU2 promoter; n = number of UASs (up to 32); hybrid	Blazeck et al., 2011
n UAS1XPR2-TEF	hybrid promoter derived from pXPR2; core minimal TEF promoter; n = number of UASs (up to 16); hybrid	Blazeck et al., 2011
pEYK1, pEYD1	strongly induced by erythritol and erythrulose; repressed by glucose and glycerol; native and hybrid (p3AB-EYK)	Trassaert et al., 2017 Park et al., unpublished
Terminators
XPRt	native terminator sequence	Franke et al., 1988
Lip2t	native terminator sequence	Pignede et al., 2000
Minimal XPRt	100-bp non-coding 3′ sequence	Swennen et al., 2002
CYC-t	S. cerevisiae terminator sequence	Blazeck et al., 2011; Mumberg et al., 1995
Synthetic	short synthetic terminators	Curran et al., 2015
Markers
LEU2, URA3, Lys5	auxotrophy complementation	Barth and Gaillardin, 1996
Ura3d4	promoter-defective gene; several copies needed to restore auxotrophy	Le Dall et al., 1994
SUC2	gene from S. cerevisiae; sucrose utilization	Nicaud et al., 1989
EYK1	gene from Y. lipolytica; erythritol as carbon source	Vandermies et al., 2017
ptxD	gene from Pseudomonas stutzeri; growth in phosphite-containing media	Shaw et al., 2016
Hph	gene from E. coli; resistance to Hygromycin B	Otero et al., 1996; Tsakraklides et al., 2018
nat1	gene from Streptomyces noursei; resistance to nourseothricin	Kretzschmar et al., 2013; Tsakraklides et al., 2018
guaB	gene from E. coli; resistance to mycophenolic acid	Wagner et al., 2018
ylAHAS W572 L	Y. lipolytica mutant resistant to chlorimuron ethyl herbicide	Wagner et al., 2018
Tags
tripeptide AKI or SKL	peroxisomal targeting signal	Xue et al., 2013
GPI anchor domains	signal for surface display; covalent bonds with cell wall β-1,6 glucans	Yue et al., 2008; Yuzbasheva et al., 2011; Moon et al., 2013
Flocculation domains	signals for surface display; non-covalent bonds with cell surface mannan chains	Yang et al., 2009
CBM	signals for surface display; non-covalent bonds with chitin	Duquesne et al., 2014
Pir	signals for surface display; covalent bonds with β-1,3 glucans	Duquesne et al., 2014
Oleosin C-t domain	targets lipid bodies	Han et al., 2013
Synthetic consensus secretory signal	MKFSAALLTAALA(S:V)AAAAA	Celińska et al., 2018
Fluorescent tag	reveals expression and localization	Bredeweg et al., 2017
Elements for exogenous DNA maintenance
1 kb homologous flanking fragments	required for DNA integration by HR	Barth and Gaillardin, 1996
ZETA elements	integration at a zeta locus in zeta-containing strains; random integration in zeta-free strains	Pignede et al., 2000
rDNA	repeated genomic sequences	Le Dall et al., 1994
ARS68/ARS18	for maintenance of autonomously replicating vectors	Matsuoka et al., 1993; Fournier et al., 1993

GPI: glycosyl phosphatidyl inositol, CBM: chitin-binding module, Pir: protein internal repeat.