Table 2 Plasmids used

p64ICL1

Parental vector for multicopy integration in Y. lipolytica

ura3d4 (mc)^a

[51]

p65ICL1

Parental vector for single copy integration in Y. lipolytica

ura3d1 (sc) ^a

[51]

p67RYL

Source for Y. lipolytica cytochrome P450 reductase (YlCPRwt)

ura3d4 (mc)^a

[22, 49]

pNMTS-CYP-OR-spe

Source for codon optimized human cytochrome P450 3A4 (3A4syn)

Weis et al., unpublished (IMBT, TU Graz)

pBdpTrcRed-3A4(192v)wtA305S

Source for codon optimized human cytochrome P450 reductase (hCPRsyn)

Weis et al., unpublished (IMBT, TU Graz)

ID7262313

Source for cDNA wild-type human cytochrome P450 reductase (hCPRwt)

BioCat, Germany

Cloning vectors:

pJet1.2

E. coli cloning vector

Fermentas, Germany

p64D-linker

Cloning and basic integrative multicopy vector

ura3d4 (mc) ^a

This work

p65D-linker

Cloning and basic integrative single copy vector, negative control

ura3d1 (sc) ^a

This work

Integrative vectors for CPR and CYP expression in Y. lipolytica:

p64D-hCPRwt-2D6syn

Multicopy coexpression of human wild-type CPR (hCPRwt), codon optimized CPR (hCPRsyn) or Y.lipolytica-own CPR (YlCPR) in combination with codon optimized CYP2D6 or codon optimized CYP3A4, respectively.

ura3d4 (mc) ^a

This work

p64D-hCPRsyn-2D6syn

p64D-hCPRwt-3A4syn

p64D-YlCPR-2D6syn

p64D-YlCPR-3A4syn

p65D-hCPRwt-2D6syn

Single copy coexpression of human wild-type CPR (hCPRwt) or Y. lipolytica-own CPR (YlCPR) in combination with codon optimized CYP2D6 or codon optimized CYP3A4, respectively.

ura3d4 (mc) ^a

This work

p65D-hCPRwt-3A4syn

p65D-YlCPR-2D6syn

p65D-YlCPR-3A4syn