Sequences
CRISPR-Cas Primers | sequence |
---|---|
588 FW | CTTGAAAACTTACGCTCATCTCTTCGACG |
589 RV | CGGACTTAAGAAAATCCAGGATTGTCTTTCC |
6006 FW | GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC |
6005 RV | GATCATTTATCTTTCACTGCGGAGAAG |
F ARO10 tar |
TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAACAATGGTTTTAGAGCTAGAAATA GCAAGTTAAAATAAGGCTAGTCCGTTATCAAC |
R ARO10 tar |
GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATATTGATCATTTATCTTTCACTGC GGAGAAGTTTCGAACGCCGAAACATGCGCA |
F AGA2 KO tar |
TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGTTTATGTTTTAGAGCTAGAAATAGCAA GTTAAAATAAGGCTAGTCCGTTATCAAC |
R AGA2 KO tar |
GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGTTTTGATCATTTATCTTTCACTGCGGAG AAGTTTCGAACGCCGAAACATGCGCA |
F AGA1 OE tar |
TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAAAAAGTTTTAGAGCTAGAAATAGCAAGTTA AAATAAGGCTAGTCCGTTATCAAC |
R AGA1 OE tar |
GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGTAAGATCATTTATCTTTCACTGCGGAGAAGT TTCGAACGCCGAAACATGCGCA |
CCasR1-RV |
CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTGAAC CTCGACTTTCTGGATCAGCC |
CCasR2-FW |
AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCTAGCAT GGAACCAAGACCCATGACG |
CCasR2-RV |
TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTGAAC CTCGACTTTCTGGATCAGCC |
CCasR3-RV |
TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCTGAACC TCGACTTTCTGGATCAGCC |
CCasR3FW |
AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCTAGCATG GAACCAAGACCCATGACG |
CCasR4-FW |
GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTAGCATGG AACCAAGACCCATGACG |
CCasR5-FW |
GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAGCATGG AACCAAGACCCATGACG |
CCasR5-RV |
TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCTGAACCTC GACTTTCTGGATCAGCC |
CCasR1-FW |
ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTAGCATG GAACCAAGACCCATGACG |
CCasR4-RV |
TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTGAAC CTCGACTTTCTGGATCAGCC |
CCasAga1-FW |
GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCA TGGAACCAAGACCCATGA |
CCasAga1-RV |
ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAATGTAC TAATTAAAAGATCAGAAAA |
Cellulosome primers | sequence |
---|---|
BGLI-fragA-fw | TAGTAATGCAGACACTTGCG |
BGLI-fragA-rv | CAACAGCGGTCTACTGAATC |
BGLI-fragB-fw | CACTAGCTCAGATTCAGTAGACC |
BGLI-fragB-rv | CGAGATGGACCGCAAG |
BGLI1-FW | TTTGGATCCGTCTAGATGAA |
BGL1-RV | AAACTCGAGTACTAGTATTC |
BGL1A-FW | TTTGGATCCGTCTAGATGAGATTTCCTTCAATTTTTAC |
BGL1A-RV | AAACTCGAGTACTAGTACTAATAAGGTAGGTGGGGTA |
CBHII1-FW | TTTGGATCCGTCTAGATGAA |
CBHII1-RV | AAACTCGAGTACTAGTAACT |
CIPA3A-FW | TTTGGATCCGTCTAGATGCA |
CIPA3A-RV | AAAGTCGACCTCGAGTACTA |
CIPA3B-FW | TTTGGATCCGTCTAGATGCAGTTACTTCGCTGTTT |
CIPA3B-RV | AAACTCGAGTACTAGTATTAGTGATGGTGATGGTGATGTTCGAAGGGCCCATTCGAAT |
CIPA3C-RV | AAAGTCGACCTCGAGTATCA |
EGII1-FW | TTTGGATCCGTCTAGATGAA |
EGII1-RV | AAACTCGAGTACTAGTAGAC |
Bio-His-EG-FW1 | TATTTGAATTCGCGGCCGCTTCTAGATGCATCATCACCATCACCATG |
Bio-His-EG-RV1 | TATTTCTGCAGCGGCCGCTACTAGTATTACTTTCTTGCGAGACACGAG |
Bio-ss1-HEGII-FW1 | TATTTGAATTCGCGGCCGCTTCTAGATGAAGGTTTTGATTGTCTTG |
Int-ss1-HEGII-FW1 | TTTGGATCCGTCTAGATGATGAAGGTTTTGATTGTCTTG |
Int-ss1-HEGII-RV1 | AAACTCGAGTACTAGTATTACTTTCTTGCGAGACACGAGC |
Sequences
CRISPR-Cas Primers | sequence |
---|---|
588 FW | CTTGAAAACTTACGCTCATCTCTTCGACG |
589 RV | CGGACTTAAGAAAATCCAGGATTGTCTTTCC |
6006 FW | GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC |
6005 RV | GATCATTTATCTTTCACTGCGGAGAAG |
F ARO10 tar |
TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAACAATGGTTTTAGAGCTAGAAATA GCAAGTTAAAATAAGGCTAGTCCGTTATCAAC |
R ARO10 tar |
GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATATTGATCATTTATCTTTCACTGC GGAGAAGTTTCGAACGCCGAAACATGCGCA |
F AGA2 KO tar |
TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGTTTATGTTTTAGAGCTAGAAATAGCAA GTTAAAATAAGGCTAGTCCGTTATCAAC |
R AGA2 KO tar |
GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGTTTTGATCATTTATCTTTCACTGCGGAG AAGTTTCGAACGCCGAAACATGCGCA |
F AGA1 OE tar |
TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAAAAAGTTTTAGAGCTAGAAATAGCAAGTTA AAATAAGGCTAGTCCGTTATCAAC |
R AGA1 OE tar |
GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGTAAGATCATTTATCTTTCACTGCGGAGAAGT TTCGAACGCCGAAACATGCGCA |
CCasR1-RV |
CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTGAAC CTCGACTTTCTGGATCAGCC |
CCasR2-FW |
AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCTAGCAT GGAACCAAGACCCATGACG |
CCasR2-RV |
TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTGAAC CTCGACTTTCTGGATCAGCC |
CCasR3-RV |
TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCTGAACC TCGACTTTCTGGATCAGCC |
CCasR3FW |
AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCTAGCATG GAACCAAGACCCATGACG |
CCasR4-FW |
GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTAGCATGG AACCAAGACCCATGACG |
CCasR5-FW |
GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAGCATGG AACCAAGACCCATGACG |
CCasR5-RV |
TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCTGAACCTC GACTTTCTGGATCAGCC |
CCasR1-FW |
ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTAGCATG GAACCAAGACCCATGACG |
CCasR4-RV |
TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTGAAC CTCGACTTTCTGGATCAGCC |
CCasAga1-FW |
GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCA TGGAACCAAGACCCATGA |
CCasAga1-RV |
ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAATGTAC TAATTAAAAGATCAGAAAA |
Cellulosome primers | sequence |
---|---|
BGLI-fragA-fw | TAGTAATGCAGACACTTGCG |
BGLI-fragA-rv | CAACAGCGGTCTACTGAATC |
BGLI-fragB-fw | CACTAGCTCAGATTCAGTAGACC |
BGLI-fragB-rv | CGAGATGGACCGCAAG |
BGLI1-FW | TTTGGATCCGTCTAGATGAA |
BGL1-RV | AAACTCGAGTACTAGTATTC |
BGL1A-FW | TTTGGATCCGTCTAGATGAGATTTCCTTCAATTTTTAC |
BGL1A-RV | AAACTCGAGTACTAGTACTAATAAGGTAGGTGGGGTA |
CBHII1-FW | TTTGGATCCGTCTAGATGAA |
CBHII1-RV | AAACTCGAGTACTAGTAACT |
CIPA3A-FW | TTTGGATCCGTCTAGATGCA |
CIPA3A-RV | AAAGTCGACCTCGAGTACTA |
CIPA3B-FW | TTTGGATCCGTCTAGATGCAGTTACTTCGCTGTTT |
CIPA3B-RV | AAACTCGAGTACTAGTATTAGTGATGGTGATGGTGATGTTCGAAGGGCCCATTCGAAT |
CIPA3C-RV | AAAGTCGACCTCGAGTATCA |
EGII1-FW | TTTGGATCCGTCTAGATGAA |
EGII1-RV | AAACTCGAGTACTAGTAGAC |
Bio-His-EG-FW1 | TATTTGAATTCGCGGCCGCTTCTAGATGCATCATCACCATCACCATG |
Bio-His-EG-RV1 | TATTTCTGCAGCGGCCGCTACTAGTATTACTTTCTTGCGAGACACGAG |
Bio-ss1-HEGII-FW1 | TATTTGAATTCGCGGCCGCTTCTAGATGAAGGTTTTGATTGTCTTG |
Int-ss1-HEGII-FW1 | TTTGGATCCGTCTAGATGATGAAGGTTTTGATTGTCTTG |
Int-ss1-HEGII-RV1 | AAACTCGAGTACTAGTATTACTTTCTTGCGAGACACGAGC |
Sequencing primers | sequence |
---|---|
genseqFW | GCAGACGACTCGAATAAG |
genseqRV | AGCTCGTCAATAGTGGAGATTAC |
VR primer fw | ATTACCGCCTTTGAGTGAGC |
VR primer rv | TGCCACCTGACGTCTAAGAA |
M13 fw | GTAAAACGACGGCCAGT |
M13 rv | CAGGAAACAGCTATGAC |
EGII-seq-fw | GGCTGTACCACAGATGG |
EGII-seq-rv | GTTCTGTCGGAGCCAAG |
BGL1-seq-paper-FW | GCAACAGCCAAGCATTAC |
BGL1-seq2-paper-FW | ACCACAGCGAGGTTATTC |
PAL2-seq-ver-RV | GTGGTGCTCATCAGTTTC |
PAL2-seq-fw-internal | CTGCGTGGCACCATTAC |
Zeocin cassette primers | sequence |
---|---|
Zcas1-FW | TATTTGAATTCGCGGCCGCTTCTAGAGTCCAGAAAGTCGAGGTTCAG |
Zcas2-FW | TATTTGAATTCGCGGCCGCTTCTAGAGGGGCTGATCCAGAAAGTCGA |
Zcas3-FW | TATTTGAATTCGCGGCCGCTTCTAGAGCCAGAAAGTCGAGGTTCAG |
Zcas4-FW | TATTTGAATTCGCGGCCGCTTCTAGAGGATCCAGAAAGTCGAGGTTCAG |
Zcas1-RV | TATTTCTGCAGCGGCCGCTACTAGTATTATGCTTCCGGCTCGTATG |
Zcas2-RV | TATTTCTGCAGCGGCCGCTACTAGTACTTCCGGCTCGTATGTTGTG |
Zcas3-RV | TATTTCTGCAGCGGCCGCTACTAGTAGCTCGGAATTAACCCTCAC |
Synthetic DNA sequences | Source |
---|---|
ss1-V5-EGII-DocS | "Ordered from IDTA, designed based on Wen et al. 2010" |
ss1-FLAG-CBHI-DocS | "Ordered from IDTA, designed based on Wen et al. 2010" |
ss1-myc-BGLI-DocA part1 | "Ordered from IDTA, designed based on Wen et al. 2010" |
ss1-myc-BGLI-DocA part2 | "Ordered from IDTA, designed based on Wen et al. 2010" |
AGA2-CIPA3-His part1 | "Ordered from IDTA, designed based on Wen et al. 2010" |
AGA2-CIPA3-His part2 | "Ordered from IDTA, designed based on Wen et al. 2010" |
synthetic PAL2 gene part1 | "Ordered from IDTA, designed based on McKenna, 2014" |
synthetic PAL2 gene part2 | "Ordered from IDTA, designed based on McKenna, 2014" |
Strains | Source |
---|---|
BJ1991 | "Molecular Cell Biology group, RUG" |
BY4741 | "Molmic group, RUG" |
BJC3CB | This work |
BJC1CB | This work |
YPH499 | "Molecular Cell Biology group, RUG" |
YPHPAL2 | This work |
YPHCON | This work |
Plasmids | Source |
---|---|
pYD1-CipA3-EGII | From Wen et al. 2010 |
pYD1-CipA1-EGII | From Wen et al. 2010 |
pRS425-CBHII-BGL1 | From Wen et al. 2010 |
p414 KanMX TEF1-Cas9-Tcyc | "Molmic group, RUG" |
pMEL16 OR1 | "Molmic group, RUG" |
pMEL16-ARO10tar | This work |
pMEL16-AGA2tar | This work |
pMEL16-AGA1tar | This work |
pUC57-PAL2 | "Roelfes group, RUG" |
pRS313+ s promHxt7-MCS-terHxt7 | "Molmic group, RUG" |
pRS313-Hxt7-PAL2 | This work |
Sequencing primers | sequence |
---|---|
genseqFW | GCAGACGACTCGAATAAG |
genseqRV | AGCTCGTCAATAGTGGAGATTAC |
VR primer fw | ATTACCGCCTTTGAGTGAGC |
VR primer rv | TGCCACCTGACGTCTAAGAA |
M13 fw | GTAAAACGACGGCCAGT |
M13 rv | CAGGAAACAGCTATGAC |
EGII-seq-fw | GGCTGTACCACAGATGG |
EGII-seq-rv | GTTCTGTCGGAGCCAAG |
BGL1-seq-paper-FW | GCAACAGCCAAGCATTAC |
BGL1-seq2-paper-FW | ACCACAGCGAGGTTATTC |
PAL2-seq-ver-RV | GTGGTGCTCATCAGTTTC |
PAL2-seq-fw-internal | CTGCGTGGCACCATTAC |
Zeocin cassette primers | sequence |
---|---|
Zcas1-FW | TATTTGAATTCGCGGCCGCTTCTAGAGTCCAGAAAGTCGAGGTTCAG |
Zcas2-FW | TATTTGAATTCGCGGCCGCTTCTAGAGGGGCTGATCCAGAAAGTCGA |
Zcas3-FW | TATTTGAATTCGCGGCCGCTTCTAGAGCCAGAAAGTCGAGGTTCAG |
Zcas4-FW | TATTTGAATTCGCGGCCGCTTCTAGAGGATCCAGAAAGTCGAGGTTCAG |
Zcas1-RV | TATTTCTGCAGCGGCCGCTACTAGTATTATGCTTCCGGCTCGTATG |
Zcas2-RV | TATTTCTGCAGCGGCCGCTACTAGTACTTCCGGCTCGTATGTTGTG |
Zcas3-RV | TATTTCTGCAGCGGCCGCTACTAGTAGCTCGGAATTAACCCTCAC |
Synthetic DNA sequences | Source |
---|---|
ss1-V5-EGII-DocS | "Ordered from IDTA, designed based on Wen et al. 2010" |
ss1-FLAG-CBHI-DocS | "Ordered from IDTA, designed based on Wen et al. 2010" |
ss1-myc-BGLI-DocA part1 | "Ordered from IDTA, designed based on Wen et al. 2010" |
ss1-myc-BGLI-DocA part2 | "Ordered from IDTA, designed based on Wen et al. 2010" |
AGA2-CIPA3-His part1 | "Ordered from IDTA, designed based on Wen et al. 2010" |
AGA2-CIPA3-His part2 | "Ordered from IDTA, designed based on Wen et al. 2010" |
synthetic PAL2 gene part1 | "Ordered from IDTA, designed based on McKenna, 2014" |
synthetic PAL2 gene part2 | "Ordered from IDTA, designed based on McKenna, 2014" |
Strains | Source |
---|---|
BJ1991 | "Molecular Cell Biology group, RUG" |
BY4741 | "Molmic group, RUG" |
BJC3CB | This work |
BJC1CB | This work |
YPH499 | "Molecular Cell Biology group, RUG" |
YPHPAL2 | This work |
YPHCON | This work |
Plasmids | Source |
---|---|
pYD1-CipA3-EGII | From Wen et al. 2010 |
pYD1-CipA1-EGII | From Wen et al. 2010 |
pRS425-CBHII-BGL1 | From Wen et al. 2010 |
p414 KanMX TEF1-Cas9-Tcyc | "Molmic group, RUG" |
pMEL16 OR1 | "Molmic group, RUG" |
pMEL16-ARO10tar | This work |
pMEL16-AGA2tar | This work |
pMEL16-AGA1tar | This work |
pUC57-PAL2 | "Roelfes group, RUG" |
pRS313+ s promHxt7-MCS-terHxt7 | "Molmic group, RUG" |
pRS313-Hxt7-PAL2 | This work |
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