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<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>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAA<br>CAATGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R ARO10 tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATA<br>TTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA2 KO tar</td><td>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGT<br>TTATGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA2 KO tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGT<br>TTTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA1 OE tar</td><td>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAA<br>AAAGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA1 OE tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGT<br>AAGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>CCasR1-RV</td><td>CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTG<br>AACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR2-FW</td><td>AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCT<br>AGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR2-RV</td><td>TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTG<br>AACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3-RV</td><td>TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCT<br>GAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3FW</td><td>AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCT<br>AGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-FW</td><td>GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTA<br>GCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR5-FW</td><td>GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAG<br>CATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR5-RV</td><td>TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCT<br>GAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR1-FW</td><td>ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTA<br>GCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-RV</td><td>TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTG<br>AACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasAga1-FW</td><td>GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCATGGAA<br>CCAAGACCCATGA</td></tr><tr><td>CCasAga1-RV</td><td>ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAAT<br>GTACTAATTAAAAGATCAGAAAA</td></tr></table> | <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>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAA<br>CAATGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R ARO10 tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATA<br>TTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA2 KO tar</td><td>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGT<br>TTATGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA2 KO tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGT<br>TTTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA1 OE tar</td><td>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAA<br>AAAGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA1 OE tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGT<br>AAGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>CCasR1-RV</td><td>CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTG<br>AACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR2-FW</td><td>AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCT<br>AGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR2-RV</td><td>TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTG<br>AACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3-RV</td><td>TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCT<br>GAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3FW</td><td>AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCT<br>AGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-FW</td><td>GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTA<br>GCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR5-FW</td><td>GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAG<br>CATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR5-RV</td><td>TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCT<br>GAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR1-FW</td><td>ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTA<br>GCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-RV</td><td>TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTG<br>AACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasAga1-FW</td><td>GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCATGGAA<br>CCAAGACCCATGA</td></tr><tr><td>CCasAga1-RV</td><td>ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAAT<br>GTACTAATTAAAAGATCAGAAAA</td></tr></table> | ||
<div class="clear"></div> | <div class="clear"></div> |
Revision as of 19:16, 17 October 2018
CRISPR-Cas Primers | sequence |
---|---|
588 FW | CTTGAAAACTTACGCTCATCTCTTCGACG |
589 RV | CGGACTTAAGAAAATCCAGGATTGTCTTTCC |
6006 FW | GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC |
6005 RV | GATCATTTATCTTTCACTGCGGAGAAG |
F ARO10 tar | TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAA CAATGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC |
R ARO10 tar | GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATA TTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA |
F AGA2 KO tar | TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGT TTATGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC |
R AGA2 KO tar | GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGT TTTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA |
F AGA1 OE tar | TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAA AAAGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC |
R AGA1 OE tar | GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGT AAGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA |
CCasR1-RV | CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTG AACCTCGACTTTCTGGATCAGCC |
CCasR2-FW | AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCT AGCATGGAACCAAGACCCATGACG |
CCasR2-RV | TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTG AACCTCGACTTTCTGGATCAGCC |
CCasR3-RV | TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCT GAACCTCGACTTTCTGGATCAGCC |
CCasR3FW | AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCT AGCATGGAACCAAGACCCATGACG |
CCasR4-FW | GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTA GCATGGAACCAAGACCCATGACG |
CCasR5-FW | GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAG CATGGAACCAAGACCCATGACG |
CCasR5-RV | TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCT GAACCTCGACTTTCTGGATCAGCC |
CCasR1-FW | ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTA GCATGGAACCAAGACCCATGACG |
CCasR4-RV | TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTG AACCTCGACTTTCTGGATCAGCC |
CCasAga1-FW | GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCATGGAA CCAAGACCCATGA |
CCasAga1-RV | ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAAT GTACTAATTAAAAGATCAGAAAA |
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 |
PAL2 primers | sequence |
---|---|
PAL2A-FW | TTTGGATCCGTCTAGATGGA |
PAL2A-RV | AAACTCGAGTACTAGTATTAGCAAATCGGAATCGGAGCTC |
PAL2B-FW | TTTGGATCCATGCATCACCATCACCATCACGATCAAATCGAAGCAATGTT |
PAL2B-RV | AAACTCGAGTACTAGTATTAATGGTGATGGTGATGGTGGCAAATCGGAATCGGAGC |
PAL2C-FW | TTTGGATCCGTCTAGATGGATCAAATCGAAGCAAT |
PAL2.1-FW | CACTAGCTCAGATTCAGTAG |
PAL2.1-RV | AAACCATGGATAGCCTTGTT |
PAL2.2-RV | TTAGCAGATTGGGATTGGGG |
PAL2D-FW | ATATACCAGCATTTAAGAACCTAATGAGTTCTGTTTGTAATGCG |
PAL2D-RV | AGGTTCTTAAATGCTGGTATATTCGGAAACACGAAGGAGA |
Pal2F-fw | TTTGGATCCGTCTAGATGGATCAAATAGAAGCAAT |
Pal2F-rv | AAACTCGAGTTAGTGATGGTGATGGTGATGGCAGATTGGGATTGGGGCAC |
Pal2G-fw | TTTGGATCCATGCATCACCATCACCATCACATGGATCAAATAGAAGCAAT |
Pal2G-rv | AAACTCGAGTACTAGTATTAGCAGATTGGGATTGGGG |
PAL2-fragA-fw | TTTGGATCCGATGGATCAAATAGAAGCAATGTTG |
PAL2-fragA-rv | AAACCATGGATAGCCTTGTTTC |
PAL2-fragB-fw | ACTAGCTCAGATTCAGTAGACC |
PAL2-fragB-rv | AAACTCGAGTTAGCAGATTGGGATTGGGG |
P T7 | GACCATCACACCACTGAAGACTGCGG |
R T7 | GTGATCATGAATTAATAAAAGTGTTCGC |
PAL2-BB-fix-fw | TTAATGAATTCGCGGCCGCTTCTAGATGGACCAAATTGAGGCGATG |
PAL2-BB-fix-rv | TTAATCTGCAGCGGCCGCTACTAGTACAGTTAGCAAATCGGAATCG |
PAL2-BB-fw | GAATTCGCGGCCGCTTCTAGATGGACCAAATTGAGGCGATG |
PAL2-BB-rv | CTGCAGCGGCCGCTACTAGTACAGTTAGCAAATCGGAATCG |
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 |