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. 2019 Jul 2;12:173. doi: 10.1186/s13068-019-1486-8

Table 4.

List of primers used in this work

Primer name Sequence Explanation
pRCC2_Fw TGTTGTCTGACATTTTGAGAGTTAACACCGAAATTACCAAGGCTC Primer for pRCCK/pRCCN amplification
pRCC1_Rv CTTGGTGGTGTTCGTCGTATCTCTTAATCATAGAAGCAGACAATGGAG Primer for pRCCK/pRCCN amplification
CC-Bdh1_Fw

GAAAATCTATGTACCCACGCGTTTTAGAGCTAGAAATAGCAAGTTAAA

ATAAGG

BDH1 target for pRCCK
CC-Bdh1_Rv GCGTGGGTACATAGATTTTCGATCATTTATCTTTCACTGCGGAG BDH1 target for pRCCK
JWP001-DR-BDH1/2

GCAATAAGAATAACAATAAATTCATTGAACATATTTCAGATGACAAAAT

AATATTTGGGGCCCCTCGCGGCTCATTTGTA

Donor DNA for ∆bdh1
JWP002-DR-BDH1/2c

TACAAATGAGCCGCGAGGGGCCCCAAATATTATTTTGTCATCTGAAAT

ATGTTCAATGAATTTATTGTTATTCTTATTGC

Donor DNA for ∆bdh1
A1-Bdh2 TGACTGTGTTTGTGGTTCTC PCR control of ∆bdh1 and ∆bdh2
A4-Bdh1 TCGTCTTTGTTCCCACATTC PCR control of ∆bdh1 and ∆bdh2
CC-Leu4-9_Fw

ACAATGACCGTGGTTGGTTTTAGAGCTAGAAATAGCAAGTTAAAATA

AGG

LEU4/9 target for pRCCK
CC-Leu4-9_Rv CAACCACGGTCATTGTGACAGATCATTTATCTTTCACTGCGGAG LEU4/9 target for pRCCK
DR-Leu4

TACTGTAGACTTTTTCCTTACAAAAAGACAAGGAACAATCGAACTTTTC

TGTATTTCAGGACTTATTCGCTTCTATTTAT

Donor DNA for ∆leu4
JWP003-DR-Leu4c

ATAAATAGAAGCGAATAAGTCCTGAAATACAGAAAAGTTCGATTGTTC

CTTGTCTTTTTGTAAGGAAAAAGTCTACAGTA

Donor DNA for ∆leu4
A1-Leu4 TTGTACAGTAACGGCCAGTC PCR control of ∆leu4
A4-Leu4 TTCGTCACTAACCGCCAAAC PCR control of ∆leu4
DR-Leu9

GGATAATACTATCGGCACATTATCATTTAGCCGCGTAGCCTAGAAAGG

AGTAGCTTATGATTACTCATGTTATATATATA

Donor DNA for ∆leu9
JWP004-DR-Leu9c

TATATATATAACATGAGTAATCATAAGCTACTCCTTTCTAGGCTACGCG

GCTAAATGATAATGTGCCGATAGTATTATCC

Donor DNA for ∆leu9
A1-Leu9 GGTAACGGTCGTAGTGAATG PCR control of ∆leu9
A4-Leu9 TGTTCTCCCTTCACAAAGTC PCR control of ∆leu9
CC-Ecm31_Fw

GAAGAACTGTGCTCCCGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAA

GG

ECM31 target for pRCCN
CC-Ecm31_Rv GGAGCACAGTTCTTCAATGATCATTTATCTTTCACTGCGGAG ECM31 target for pRCCN
DR-Ecm31

ATTAGCTTGCCATAAAATTAGGGAAATTTTTACTCACAATAATATATAGA

TAAAAATCACTGCATAGGGAAAAAAACTTT

Donor DNA for ∆ecm31
JWP005_DR-Ecm31c

AAAGTTTTTTTCCCTATGCAGTGATTTTTATCTATATATTATTGTGAGTA

AAAATTTCCCTAATTTTATGGCAAGCTAAT

Donor DNA for ∆ecm31
A1-Ecm31 ATGTACACGACAGACATTCC PCR control of ∆ecm31
A4-Ecm31 TATTATAAAGCGGCCAGCTC PCR control of ∆ecm31
CC-Ilv1_Fw

TACTTTACCCGACGTCCCGTTTTAGAGCTAGAAATAGCAAGTTAAAAT

AAGG

ILV1 target for pRCCK
CC-Ilv1_Rv GACGTCGGGTAAAGTAACGATCATTTATCTTTCACTGCGGAG ILV1 target for pRCCK
DR-Ilv1

CAAGCCACATTTAAACTAAGTCAATTACACAAAGTTAGTGAACCGACA

ATTTACTTTATAAATTTACGCAACAACTTGTT

Donor DNA for ∆ilv1
JWP006-DR-ilv1c

AACAAGTTGTTGCGTAAATTTATAAAGTAAATTGTCGGTTCACTAACT

TTGTGTAATTGACTTAGTTTAAATGTGGCTTG

Donor DNA for ∆ilv1
A1-Ilv1 AATTCACTAGCGGCTCCTTG PCR control of ∆ilv1
A4-Ilv1 ATGGCTATGTGGAAGAAGTC PCR control of ∆ilv1
CC-Bat12_Fw

ACAAGAGCTTGGCCAGGGTTTTAGAGCTAGAAATAGCAAGTTAAAA

TAAGG

BAT1 and BAT2 target for pRCCK
CC-Bat1_Rv CCTGGCCAAGCTCTTGTAGCGATCATTTATCTTTCACTGCGGAG BAT1 target for pRCCK
CC-Bat2_Rv CCTGGCCAAGCTCTTGTGGCGATCATTTATCTTTCACTGCGGAG BAT2 target for pRCCK
DR-Bat1

TATAAACGCAAAATCAGCTAGAACCTTAGCATACTAAAACTGATAA

TGAAGGTAAACATCCCCTCCCCCCCCAAAAAAAA

Donor DNA for ∆bat1
JWP007-DR-Bat1c

TTTTTTTTGGGGGGGGAGGGGATGTTTACCTTCATTATCAGTTTTAG

TATGCTAAGGTTCTAGCTGATTTTGCGTTTATA

Donor DNA for ∆bat1
A1-Bat1 TTTAATGGCCCATCCGATCC PCR control of ∆bat1
A4-Bat1 AAGTCCAGCGAGATACCTTG PCR control of ∆bat1
DR-Bat2

AAATTTAAGGGAAAGCATCTCCACGAGTTTTAAGAACGATAGTATCG

CTATTGCTACGTAAAGTAATTAAAAGTTAAAAA

Donor DNA for ∆bat2
JWP008-DR-Bat2c

TTTTTAACTTTTAATTACTTTACGTAGCAATAGCGATACTATCGTTCT

TAAAACTCGTGGAGATGCTTTCCCTTAAATTT

Donor DNA for ∆bat2
A1-Bat2 GTGAGAGGAGATCCGAAATGAG PCR control of ∆bat2
A4-Bat2 TCCACCGACATTACGGAAAC PCR control of ∆bat2
JWP051-CC-Pdc1_Fw(4)

TGTTCCAGACACGACGTCAGTTTTAGAGCTAGAAATAGCAAGTTAA

AATAAGG

PDC1 target for pRCCK
JWP052-CC-Pdc1_Rv(4) CTGACGTCGTGTCTGGAACAGATCATTTATCTTTCACTGCGGAG PDC1 target for pRCCK
DR_PDC1-Fw

TCTCAATTATTATCTTCTACTCATAACCTCACGCAAAATAACACAGT

CAAATCAATCAAAGCGATTTAATCTCTAATTATTAGTTAAAGTTTTA

TAAGCATTTTTATGTAACGAAAAATA

Donor DNA for ∆pdc1
DR_PDC1-Rv

TATTTTTCGTTACATAAAAATGCTTATAAAACTTTAACTAATAATTA

GAGATTAAATCGCTTTGATTGATTTGACTGTGTTATTTTGCGTGAG

GTTATGAGTAGAAGATAATAATTGAGA

Donor DNA for ∆pdc1
A1-PDC1 GAAATCAGCTTGTGGGTATTGTTCAGAG PCR control of ∆pdc1
A4-PDC1 CCTGGTGGCATTTGCAAAATG PCR control of ∆pdc1
JWP021-CC-Pdc5_Fw

ACGAAGTAACCTCACAATCGTTTTAGAGCTAGAAATAGCAAGT

TAAAATAAGG

PDC5 target for pRCCK
JWP022-CC-Pdc5_Rv CGATTGTGAGGTTACTTCGTGATCATTTATCTTTCACTGCGGAG PDC5 target for pRCCK
DR_PDC5-Fw

ACTTATTTCACATAATCAATCTCAAAGAGAACAACACAATACAATAA

CAAGAAGAACAAAGCTAATTAACATAAAACTCATGATTCAACGTTT

GTGTATTTTTTTACTTTTGAAGGTTAT

Donor DNA for ∆pdc5
DR_PDC5-Rv

ATAACCTTCAAAAGTAAAAAAATACACAAACGTTGAATCATGAGTTT

TATGTTAATTAGCTTTGTTCTTCTTGTTATTGTATTGTGTTGTTCTCTT

TGAGATTGATTATGTGAAATAAGT

Donor DNA for ∆pdc5
A1-PDC5 CGTATACGAATTCCTTCAACAAAGGCC PCR control of ∆pdc5
A4-PDC5 TAAGAAGGCATGTTGGCCTCTGTTTC PCR control of ∆pdc5
JWP023-Mth1/Pdc1_Fw

TGCTTATAAAACTTTAACTAATAATTAGAGATTAAATCGCATGTTTG

TTTCACCACCACCAG

Primer for synthesis of Donor DNA of Mth1 wt
JWP024-Mth1/Pdc1_Rv

TCATAACCTCACGCAAAATAACACAGTCAAATCAATCAAATCAGGA

TACTGAATCCGGCTGC

Primer for synthesis of Donor DNA of Mth1 wt
JWP025-Mth1-∆T_Rv

CATTAGTTAGTTGCGTGTGCACAGTAGAGGGGGCAGAAAACATTG

ATAGTGGCAAACTTTG

Primer for synthesis of Donor DNA of Mth1∆T
JWP026-Mth1-∆T_Fw

CAGTGATAATGCTTCTTTTCAAAGTTTGCCACTATCAATGTTTTCTGC

CCCCTCTACTGTG

Primer for synthesis of Donor DNA of Mth1∆T
JWP043-CC-MTH1_Fw

GCAGTATGCATTCAGCGAGCGTTTTAGAGCTAGAAATAGCAAGT

TAAAATAAGG

MTH1 target for pRCCK
JWP044-CC-MTH1_Rv CGCTCGCTGAATGCATACTGCGATCATTTATCTTTCACTGCGGAG MTH1 target for pRCCK
JWP045-DR-MTH1dT_Fw

CAGTGATAATGCTTCTTTTCAAAGTTTGCCACTATCAATGTTTTCTGCCCC

CTCTACTGTGCACACGCAACTAACTAATG

Donor DNA for MTH1∆T modification
JWP046-DR-MTH1dT_Rv

CATTAGTTAGTTGCGTGTGCACAGTAGAGGGGGCAGAAAACATTGATAG

TGGCAAACTTTGAAAAGAAGCATTATCACTG

Donor DNA for MTH1∆T modification
JWP047-MTH1_Fw ATGTTTGTTTCACCACCACCAGC PCR control of MTH1 modifications
JWP048-MTH1_Rv TCAGGATACTGAATCCGGCTGC PCR control of MTH1 modifications
JWP053-CC-Adh1_Fw

TAACTTGATGGCCGGTCACTGTTTTAGAGCTAGAAATAGCAAGTT

AAAATAAGG

ADH1 target for pRCCK
JWP054-CC-Adh1_Rv CAGTGACCGGCCATCAAGTTAGATCATTTATCTTTCACTGCGGAG ADH1 target for pRCCK
JWP055-DR-Adh1_Fw

TCAAGCTATACCAAGCATACAATCAACTATCTCATATACAGCGAAT

TTCTTATGATTTATGATTTTTATTATTAAATAAG

Donor DNA for ∆adh1
JWP056-DR-Adh1_Rv

CTTATTTAATAATAAAAATCATAAATCATAAGAAATTCGCTGTATA

TGAGATAGTTGATTGTATGCTTGGTATAGCTTGA

Donor DNA for ∆adh1
Sbp26-Chk_DelADH1_A1 GCAACCAAACCCATACATCG PCR control of ∆adh1
Sbp27-Chk_DelADH1_A4 GGGCGGAGCGTTCTAATTG PCR control of ∆adh1
vsp381_CC-GPD1_rev GCCTAGACCTTCGACGAAACGATCATTTATCTTTCACTGCGGAG GPD1 target for pRCCK
vsp382_CC-GPD1_fw

GTTTCGTCGAAGGTCTAGGCGTTTTAGAGCTAGAAATAGCAAGTT

AAAATAAGG

GPD1 target for pRCCK
vsp383_CC-GPD1-Donor

CACGTAGACTGGCTTGGTATTGGCAGTTTCGTAGTTATATATTTAT

TGGAGAAAGATAACATATCATACTTTCCCCCACT

Donor DNA for ∆gpd1
JWP058-DR-Gpd1_Rv

AGTGGGGGAAAGTATGATATGTTATCTTTCTCCAATAAATATATA

ACTACGAAACTGCCAATACCAAGCCAGTCTACGTG

Donor DNA for ∆gpd1
vsp384_pGPD1_fw GTACAGCTGATGGGACCTTGCCG PCR control of ∆gpd1
vsp385_tGPD1_rev GCTCCGTATTATCTTCGTCGTGGGG PCR control of ∆gpd1
JWP059-CC-Gpd2_Fw

CCCTTACATGAGGGGCCACGGTTTTAGAGCTAGAAATAGCAAGTT

AAAATAAGG

GPD2 target for pRCCK
JWP060-CC-Gpd2_Rv CCGTGGCCCCTCATGTAAGGGGATCATTTATCTTTCACTGCGGAG GPD2 target for pRCCK
JWP061-DR-Gpd2_Fw

CTCTTTCCCTTTCCTTTTCCTTCGCTCCCCTTCCTTATCAACACTCTCC

CCCCCCCTCCCCCTCTGATCTTTCCTGTTGC

Donor DNA for ∆gpd2
JWP062-DR-Gpd2_Rv

GCAACAGGAAAGATCAGAGGGGGAGGGGGGGGGAGAGTGTTGA

TAAGGAAGGGGAGCGAAGGAAAAGGAAAGGGAAAGAG

Donor DNA for ∆gpd2
vsp269_pGPD2_fw GGAACATCCGAGCACCCGCGCC PCR control of ∆gpd2
vsp270_tGPD2_rev GGCGGCATCGAAATCTTCTTCTTGCCC PCR control of ∆gpd2
Vsp388_CC-ALD6_fw AAAACTTTGGCCTTAGCCCGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGG ALD6 target for pRCCK
vsp389_CC-ALD6_rev CGGGCTAAGGCCAAAGTTTTGATCATTTATCTTTCACTGCGGAG ALD6 target for pRCCK
vsp390_CC-ALD6-Donor AACATCTTTAACATACACAAACACATACTATCAGAATACATGTACCAACCTGCATTTCTTTCCGTCATATACACAAAATA Donor DNA for ∆ald6
JWY092_DR_Ald6_c TATTTTGTGTATATGACGGAAAGAAATGCAGGTTGGTACATGTATTCTGATAGTATGTGTTTGTGTATGTTAAAGATGTT Donor DNA for ∆ald6
vsp393_ALD6_rev TACCGGCCTTCAACATCTTGGCC PCR control of ∆ald6
vsp394_ALD6_fw TCCACGACACTGAATGGGCTACCC PCR control of ∆ald6