Table 1 Table of the oligonucleotides used. These include the bridge primers for adding the sgRNA sequences to the pKOPIS + sgRNA plasmid, the primers for PCR SOEing to clone the donor DNA, and the primers for PCR and Sanger sequencing
NameSequence
ADE2 Bridge L1cgggtggcgaatgggactttACAGTTGGTATATTAGGAGGgttttagagctagaaatagc
ADE2 Bridge L2cgggtggcgaatgggactttAACAGTTGGTATATTAGGAGgttttagagctagaaatagc
ADE2 Bridge H1cgggtggcgaatgggactttACTTTGGCATACGATGGAAGgttttagagctagaaatagc
ADE2 Bridge H2cgggtggcgaatgggactttACGGAGTCCGGAACTCTAGCgttttagagctagaaatagc
ADE2 5′ KO ForgatgtccacgacgtctctCAAATGACTCTTGTTGCATGG
ADE2 5′ KO RevGTATATCAATAAACTTATATAACTTGATTGTTTTGTCCGATTTTC
ADE2 3′ KO ForGAAAATCGGACAAAACAATCAAGTTATATAAGTTTATTGATATAC
ADE2 3′ KO RevcggtgtcggtgtcgtagGTATAATAAGTGATCTTATGTATG
ADE2 Conf ForACCAACATAACACTGACATC
ADE2 Conf RevTATATGAACTGTATCGAAAC
pKOPIS sgRNA ForAACGCGAGCTGCGCACATAC
pKOPIS sgRNA RevGCGACAGTCACATCATGCC
pKOPIS sgRNA Seq ForCACCTATATCTGCGTGTTG
pKOPIS sgRNA Seq RevGCACGTCAAGACTGTCAAGG