Difference between revisions of "Template:Groningen/Primers"
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TAGTAATGCAGACACTTGCG</td></tr><tr><td>BGLI-fragA-rv</td><td>CAACAGCGGTCTACTGAATC</td></tr><tr><td>BGLI-fragB-fw</td><td>CACTAGCTCAGATTCAGTAGACC</td></tr><tr><td>BGLI-fragB-rv</td><td>CGAGATGGACCGCAAG</td></tr><tr><td>BGLI1-FW</td><td>TTTGGATCCGTCTAGATGAA</td></tr><tr><td>BGL1-RV</td><td>AAACTCGAGTACTAGTATTC</td></tr><tr><td>BGL1A-FW</td><td>TTTGGATCCGTCTAGATGAGATTTCCTTCAATTTTTAC</td></tr><tr><td>BGL1A-RV</td><td>AAACTCGAGTACTAGTACTAATAAGGTAGGTGGGGTA</td></tr><tr><td>CBHII1-FW</td><td>TTTGGATCCGTCTAGATGAA</td></tr><tr><td>CBHII1-RV</td><td>AAACTCGAGTACTAGTAACT</td></tr><tr><td>CIPA3A-FW</td><td>TTTGGATCCGTCTAGATGCA</td></tr><tr><td>CIPA3A-RV</td><td>AAAGTCGACCTCGAGTACTA</td></tr><tr><td>CIPA3B-FW</td><td>TTTGGATCCGTCTAGATGCAGTTACTTCGCTGTTT</td></tr><tr><td>CIPA3B-RV</td><td>AAACTCGAGTACTAGTATTAGTGATGGTGATGGTGATGTTCGAAGGGCCCATTCGAAT</td></tr><tr><td>CIPA3C-RV</td><td>AAAGTCGACCTCGAGTATCA</td></tr><tr><td>EGII1-FW</td><td>TTTGGATCCGTCTAGATGAA</td></tr><tr><td>EGII1-RV</td><td>AAACTCGAGTACTAGTAGAC</td></tr><tr><td>Bio-His-EG-FW1</td><td>TATTTGAATTCGCGGCCGCTTCTAGATGCATCATCACCATCACCATG</td></tr><tr><td>Bio-His-EG-RV1</td><td>TATTTCTGCAGCGGCCGCTACTAGTATTACTTTCTTGCGAGACACGAG</td></tr><tr><td>Bio-ss1-HEGII-FW1</td><td>TATTTGAATTCGCGGCCGCTTCTAGATGAAGGTTTTGATTGTCTTG</td></tr><tr><td>Int-ss1-HEGII-FW1</td><td>TTTGGATCCGTCTAGATGATGAAGGTTTTGATTGTCTTG</td></tr><tr><td>Int-ss1-HEGII-RV1</td><td>AAACTCGAGTACTAGTATTACTTTCTTGCGAGACACGAGC</td></tr></table> | TAGTAATGCAGACACTTGCG</td></tr><tr><td>BGLI-fragA-rv</td><td>CAACAGCGGTCTACTGAATC</td></tr><tr><td>BGLI-fragB-fw</td><td>CACTAGCTCAGATTCAGTAGACC</td></tr><tr><td>BGLI-fragB-rv</td><td>CGAGATGGACCGCAAG</td></tr><tr><td>BGLI1-FW</td><td>TTTGGATCCGTCTAGATGAA</td></tr><tr><td>BGL1-RV</td><td>AAACTCGAGTACTAGTATTC</td></tr><tr><td>BGL1A-FW</td><td>TTTGGATCCGTCTAGATGAGATTTCCTTCAATTTTTAC</td></tr><tr><td>BGL1A-RV</td><td>AAACTCGAGTACTAGTACTAATAAGGTAGGTGGGGTA</td></tr><tr><td>CBHII1-FW</td><td>TTTGGATCCGTCTAGATGAA</td></tr><tr><td>CBHII1-RV</td><td>AAACTCGAGTACTAGTAACT</td></tr><tr><td>CIPA3A-FW</td><td>TTTGGATCCGTCTAGATGCA</td></tr><tr><td>CIPA3A-RV</td><td>AAAGTCGACCTCGAGTACTA</td></tr><tr><td>CIPA3B-FW</td><td>TTTGGATCCGTCTAGATGCAGTTACTTCGCTGTTT</td></tr><tr><td>CIPA3B-RV</td><td>AAACTCGAGTACTAGTATTAGTGATGGTGATGGTGATGTTCGAAGGGCCCATTCGAAT</td></tr><tr><td>CIPA3C-RV</td><td>AAAGTCGACCTCGAGTATCA</td></tr><tr><td>EGII1-FW</td><td>TTTGGATCCGTCTAGATGAA</td></tr><tr><td>EGII1-RV</td><td>AAACTCGAGTACTAGTAGAC</td></tr><tr><td>Bio-His-EG-FW1</td><td>TATTTGAATTCGCGGCCGCTTCTAGATGCATCATCACCATCACCATG</td></tr><tr><td>Bio-His-EG-RV1</td><td>TATTTCTGCAGCGGCCGCTACTAGTATTACTTTCTTGCGAGACACGAG</td></tr><tr><td>Bio-ss1-HEGII-FW1</td><td>TATTTGAATTCGCGGCCGCTTCTAGATGAAGGTTTTGATTGTCTTG</td></tr><tr><td>Int-ss1-HEGII-FW1</td><td>TTTGGATCCGTCTAGATGATGAAGGTTTTGATTGTCTTG</td></tr><tr><td>Int-ss1-HEGII-RV1</td><td>AAACTCGAGTACTAGTATTACTTTCTTGCGAGACACGAGC</td></tr></table> | ||
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| + | <h1>Sequences</h1> | ||
| + | <table><tr><th>CRISPR-Cas Primers</th><th>sequence</th></tr><tr><td>588 FW</td><td> | ||
| + | CTTGAAAACTTACGCTCATCTCTTCGACG</td></tr><tr><td>589 RV</td><td>CGGACTTAAGAAAATCCAGGATTGTCTTTCC</td></tr><tr><td>6006 FW</td><td> | ||
| + | GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC</td></tr><tr><td>6005 RV</td><td>GATCATTTATCTTTCACTGCGGAGAAG</td></tr><tr><td>F ARO10 tar</td><td> | ||
| + | TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAACAATGGTTTTAGAGCTAGAAATA<br />GCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R ARO10 tar</td><td> | ||
| + | GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATATTGATCATTTATCTTTCACTGC<br />GGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA2 KO tar</td><td> | ||
| + | TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGTTTATGTTTTAGAGCTAGAAATAGCAA<br />GTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA2 KO tar</td><td> | ||
| + | GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGTTTTGATCATTTATCTTTCACTGCGGAG<br />AAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA1 OE tar</td><td> | ||
| + | TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAAAAAGTTTTAGAGCTAGAAATAGCAAGTTA<br />AAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA1 OE tar</td><td> | ||
| + | GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGTAAGATCATTTATCTTTCACTGCGGAGAAGT<br />TTCGAACGCCGAAACATGCGCA</td></tr><tr><td>CCasR1-RV</td><td> | ||
| + | CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTGAAC<br />CTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR2-FW</td><td> | ||
| + | AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCTAGCAT<br />GGAACCAAGACCCATGACG</td></tr><tr><td>CCasR2-RV</td><td> | ||
| + | TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTGAAC<br />CTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3-RV</td><td> | ||
| + | TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCTGAACC<br />TCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3FW</td><td> | ||
| + | AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCTAGCATG<br />GAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-FW</td><td> | ||
| + | GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTAGCATGG<br />AACCAAGACCCATGACG</td></tr><tr><td>CCasR5-FW</td><td> | ||
| + | GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAGCATGG<br />AACCAAGACCCATGACG</td></tr><tr><td>CCasR5-RV</td><td> | ||
| + | TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCTGAACCTC<br />GACTTTCTGGATCAGCC</td></tr><tr><td>CCasR1-FW</td><td> | ||
| + | ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTAGCATG<br />GAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-RV</td><td> | ||
| + | TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTGAAC<br />CTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasAga1-FW</td><td> | ||
| + | GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCA<br />TGGAACCAAGACCCATGA</td></tr><tr><td>CCasAga1-RV</td><td> | ||
| + | ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAATGTAC<br />TAATTAAAAGATCAGAAAA</td></tr></table> | ||
<div class="clear"></div> | <div class="clear"></div> | ||
| + | <br /> | ||
| + | <table><tr><th>Cellulosome primers</th><th>sequence</th></tr><tr><td>BGLI-fragA-fw</td><td> | ||
| + | TAGTAATGCAGACACTTGCG</td></tr><tr><td>BGLI-fragA-rv</td><td>CAACAGCGGTCTACTGAATC</td></tr><tr><td>BGLI-fragB-fw</td><td>CACTAGCTCAGATTCAGTAGACC</td></tr><tr><td>BGLI-fragB-rv</td><td>CGAGATGGACCGCAAG</td></tr><tr><td>BGLI1-FW</td><td>TTTGGATCCGTCTAGATGAA</td></tr><tr><td>BGL1-RV</td><td>AAACTCGAGTACTAGTATTC</td></tr><tr><td>BGL1A-FW</td><td>TTTGGATCCGTCTAGATGAGATTTCCTTCAATTTTTAC</td></tr><tr><td>BGL1A-RV</td><td>AAACTCGAGTACTAGTACTAATAAGGTAGGTGGGGTA</td></tr><tr><td>CBHII1-FW</td><td>TTTGGATCCGTCTAGATGAA</td></tr><tr><td>CBHII1-RV</td><td>AAACTCGAGTACTAGTAACT</td></tr><tr><td>CIPA3A-FW</td><td>TTTGGATCCGTCTAGATGCA</td></tr><tr><td>CIPA3A-RV</td><td>AAAGTCGACCTCGAGTACTA</td></tr><tr><td>CIPA3B-FW</td><td>TTTGGATCCGTCTAGATGCAGTTACTTCGCTGTTT</td></tr><tr><td>CIPA3B-RV</td><td>AAACTCGAGTACTAGTATTAGTGATGGTGATGGTGATGTTCGAAGGGCCCATTCGAAT</td></tr><tr><td>CIPA3C-RV</td><td>AAAGTCGACCTCGAGTATCA</td></tr><tr><td>EGII1-FW</td><td>TTTGGATCCGTCTAGATGAA</td></tr><tr><td>EGII1-RV</td><td>AAACTCGAGTACTAGTAGAC</td></tr><tr><td>Bio-His-EG-FW1</td><td>TATTTGAATTCGCGGCCGCTTCTAGATGCATCATCACCATCACCATG</td></tr><tr><td>Bio-His-EG-RV1</td><td>TATTTCTGCAGCGGCCGCTACTAGTATTACTTTCTTGCGAGACACGAG</td></tr><tr><td>Bio-ss1-HEGII-FW1</td><td>TATTTGAATTCGCGGCCGCTTCTAGATGAAGGTTTTGATTGTCTTG</td></tr><tr><td>Int-ss1-HEGII-FW1</td><td>TTTGGATCCGTCTAGATGATGAAGGTTTTGATTGTCTTG</td></tr><tr><td>Int-ss1-HEGII-RV1</td><td>AAACTCGAGTACTAGTATTACTTTCTTGCGAGACACGAGC</td></tr></table> | ||
| + | |||
| + | <div class="clear"></div> | ||
| + | <br /> | ||
| + | <table><tr><th>Sequencing primers</th><th>sequence</th></tr><tr><td>genseqFW</td><td>GCAGACGACTCGAATAAG</td></tr><tr><td>genseqRV</td><td>AGCTCGTCAATAGTGGAGATTAC</td></tr><tr><td>VR primer fw</td><td> | ||
| + | ATTACCGCCTTTGAGTGAGC</td></tr><tr><td>VR primer rv</td><td>TGCCACCTGACGTCTAAGAA</td></tr><tr><td>M13 fw</td><td>GTAAAACGACGGCCAGT</td></tr><tr><td>M13 rv</td><td>CAGGAAACAGCTATGAC</td></tr><tr><td> | ||
| + | EGII-seq-fw</td><td>GGCTGTACCACAGATGG</td></tr><tr><td>EGII-seq-rv</td><td>GTTCTGTCGGAGCCAAG</td></tr><tr><td>BGL1-seq-paper-FW</td><td>GCAACAGCCAAGCATTAC</td></tr><tr><td>BGL1-seq2-paper-FW</td><td> | ||
| + | ACCACAGCGAGGTTATTC</td></tr><tr><td>PAL2-seq-ver-RV</td><td>GTGGTGCTCATCAGTTTC</td></tr><tr><td>PAL2-seq-fw-internal</td><td>CTGCGTGGCACCATTAC</td></tr></table> | ||
| + | |||
| + | <div class="clear"></div> | ||
| + | <br /> | ||
| + | <table><tr><th>Zeocin cassette primers</th><th>sequence</th></tr><tr><td>Zcas1-FW</td><td>TATTTGAATTCGCGGCCGCTTCTAGAGTCCAGAAAGTCGAGGTTCAG</td></tr><tr><td>Zcas2-FW</td><td> | ||
| + | TATTTGAATTCGCGGCCGCTTCTAGAGGGGCTGATCCAGAAAGTCGA</td></tr><tr><td>Zcas3-FW</td><td>TATTTGAATTCGCGGCCGCTTCTAGAGCCAGAAAGTCGAGGTTCAG</td></tr><tr><td>Zcas4-FW</td><td> | ||
| + | TATTTGAATTCGCGGCCGCTTCTAGAGGATCCAGAAAGTCGAGGTTCAG</td></tr><tr><td>Zcas1-RV</td><td>TATTTCTGCAGCGGCCGCTACTAGTATTATGCTTCCGGCTCGTATG</td></tr><tr><td>Zcas2-RV</td><td> | ||
| + | TATTTCTGCAGCGGCCGCTACTAGTACTTCCGGCTCGTATGTTGTG</td></tr><tr><td>Zcas3-RV</td><td>TATTTCTGCAGCGGCCGCTACTAGTAGCTCGGAATTAACCCTCAC</td></tr></table> | ||
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| + | <div class="clear"></div> | ||
| + | <br /> | ||
| + | <table><tr><th>Synthetic DNA sequences</th><th>Source</th></tr><tr><td>ss1-V5-EGII-DocS</td><td>"Ordered from IDTA, designed based on Wen et al. 2010"</td></tr><tr><td>ss1-FLAG-CBHI-DocS</td><td>"Ordered from IDTA, designed based on Wen et al. 2010"</td></tr><tr><td>ss1-myc-BGLI-DocA part1</td><td>"Ordered from IDTA, designed based on Wen et al. 2010"</td></tr><tr><td>ss1-myc-BGLI-DocA part2</td><td>"Ordered from IDTA, designed based on Wen et al. 2010"</td></tr><tr><td>AGA2-CIPA3-His part1</td><td>"Ordered from IDTA, designed based on Wen et al. 2010"</td></tr><tr><td>AGA2-CIPA3-His part2</td><td>"Ordered from IDTA, designed based on Wen et al. 2010"</td></tr><tr><td>synthetic PAL2 gene part1</td><td>"Ordered from IDTA, designed based on McKenna, 2014"</td></tr><tr><td>synthetic PAL2 gene part2</td><td>"Ordered from IDTA, designed based on McKenna, 2014"</td></tr></table> | ||
| + | |||
| + | <div class="clear"></div> | ||
| + | <br /> | ||
| + | <table><tr><th>Strains</th><th>Source</th></tr><tr><td>BJ1991</td><td>"Molecular Cell Biology group, RUG"</td></tr><tr><td>BY4741</td><td>"Molmic group, RUG"</td></tr><tr><td>BJC3CB</td><td>This work</td></tr><tr><td>BJC1CB</td><td>This work</td></tr><tr><td>YPH499</td><td>"Molecular Cell Biology group, RUG"</td></tr><tr><td>YPHPAL2</td><td>This work</td></tr><tr><td>YPHCON</td><td>This work</td></tr></table> | ||
| + | |||
| + | <div class="clear"></div> | ||
| + | <br /> | ||
| + | <table><tr><th>Plasmids</th><th>Source</th></tr><tr><td>pYD1-CipA3-EGII</td><td>From Wen et al. 2010</td></tr><tr><td>pYD1-CipA1-EGII</td><td>From Wen et al. 2010</td></tr><tr><td>pRS425-CBHII-BGL1</td><td>From Wen et al. 2010</td></tr><tr><td>p414 KanMX TEF1-Cas9-Tcyc</td><td>"Molmic group, RUG"</td></tr><tr><td>pMEL16 OR1</td><td>"Molmic group, RUG"</td></tr><tr><td>pMEL16-ARO10tar</td><td>This work</td></tr><tr><td>pMEL16-AGA2tar</td><td>This work</td></tr><tr><td>pMEL16-AGA1tar</td><td>This work</td></tr><tr><td>pUC57-PAL2</td><td>"Roelfes group, RUG"</td></tr><tr><td>pRS313+ s promHxt7-MCS-terHxt7</td><td>"Molmic group, RUG"</td></tr><tr><td>pRS313-Hxt7-PAL2</td><td>This work</td></tr></table> | ||
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| + | <!--References --> | ||
| + | <!-- h4>References</h4 --> | ||
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| + | <!-- EINDE CONTENT ---------------------------------------------------> | ||
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<table><tr><th>Sequencing primers</th><th>sequence</th></tr><tr><td>genseqFW</td><td>GCAGACGACTCGAATAAG</td></tr><tr><td>genseqRV</td><td>AGCTCGTCAATAGTGGAGATTAC</td></tr><tr><td>VR primer fw</td><td> | <table><tr><th>Sequencing primers</th><th>sequence</th></tr><tr><td>genseqFW</td><td>GCAGACGACTCGAATAAG</td></tr><tr><td>genseqRV</td><td>AGCTCGTCAATAGTGGAGATTAC</td></tr><tr><td>VR primer fw</td><td> | ||
Revision as of 10:24, 15 October 2018
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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