Plasmids Description Source
pRS426 Used for cloning of pMB1 origin, AmpR, and S. cerevisiae elements of 2µ origin and URA3 Laboratory stock
pGI2 KanR for bacteria, NatR for yeast, binary plasmid (Abbott et al., 2013)
pGI2_880_ACC Used for cloning of ACC1 gene from R. toruloides (Zhang et al., 2016)
pGI2-TEF1-X Expression vector in R. toruloides, containing NatR, pTEF1 promoter and T35S terminator This study
pRTN Expression vector in R. toruloides, containing NatR, pANT promoter and T35S terminator Laboratory stock
pRTG2-X Derived from pRTN, containing G418R, pANT promoter and T35S terminator This study
pRTHyg-X Derived from pRTN, containing HygR, pANT promoter and T35S terminator This study
pRTH gRNA expression vector in R. toruloides, containing HygR, 5S-tRNATyr, 2 BsaI sites, and SUP4 terminator for SgRNA Laboratory stock
pRTH-X Derived from pRTH with target N20 insertion at BsaI sites (no BsaI left) This study
pRT-Cas9-SgRNA-Hyg Derived from pRTH-X with Cas9 expression cassette This study
Strains Description Source
DH5α Cloning host NEB
IFO0880 R. toruloides strain IFO0880, mating type A2 NBRC, Japan
I12 IFO0880 with genome integrated codon optimized GhPS cassette This study
I12-Cas9 I12 with genome integrated codon optimized Cas9 cassette This study
I12-ACL1 I12 with genome integrated ACL1 cassette This study
I12-ACL1-ACC1 I12-ACL1 with genome integrated ACC1 cassette This study