Line 17: | Line 17: | ||
<div class="column"> | <div class="column"> | ||
<h1>Sequences</h1> | <h1>Sequences</h1> | ||
+ | |||
+ | <div class="column"> | ||
<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>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAACAATGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R ARO10 tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATATTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA2 KO tar</td><td>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGTTTATGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA2 KO tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGTTTTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA1 OE tar</td><td>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAAAAAGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA1 OE tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGTAAGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>CCasR1-RV</td><td>CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR2-FW</td><td>AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR2-RV</td><td>TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3-RV</td><td>TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3FW</td><td>AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-FW</td><td>GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR5-FW</td><td>GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR5-RV</td><td>TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR1-FW</td><td>ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-RV</td><td>TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasAga1-FW</td><td>GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCATGGAACCAAGACCCATGA</td></tr><tr><td>CCasAga1-RV</td><td>ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAATGTACTAATTAAAAGATCAGAAAA</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>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAACAATGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R ARO10 tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATATTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA2 KO tar</td><td>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGTTTATGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA2 KO tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGTTTTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>F AGA1 OE tar</td><td>TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAAAAAGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC</td></tr><tr><td>R AGA1 OE tar</td><td>GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGTAAGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA</td></tr><tr><td>CCasR1-RV</td><td>CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR2-FW</td><td>AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR2-RV</td><td>TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3-RV</td><td>TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR3FW</td><td>AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-FW</td><td>GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR5-FW</td><td>GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR5-RV</td><td>TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasR1-FW</td><td>ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTAGCATGGAACCAAGACCCATGACG</td></tr><tr><td>CCasR4-RV</td><td>TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTGAACCTCGACTTTCTGGATCAGCC</td></tr><tr><td>CCasAga1-FW</td><td>GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCATGGAACCAAGACCCATGA</td></tr><tr><td>CCasAga1-RV</td><td>ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAATGTACTAATTAAAAGATCAGAAAA</td></tr></table> | ||
<div class="clear"></div> | <div class="clear"></div> | ||
Line 42: | Line 44: | ||
<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> | <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> | ||
+ | </div> | ||
<div class="clear"></div> | <div class="clear"></div> | ||
Revision as of 15:41, 14 October 2018
Sequences
CRISPR-Cas Primers | sequence |
---|---|
588 FW | CTTGAAAACTTACGCTCATCTCTTCGACG |
589 RV | CGGACTTAAGAAAATCCAGGATTGTCTTTCC |
6006 FW | GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC |
6005 RV | GATCATTTATCTTTCACTGCGGAGAAG |
F ARO10 tar | TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAACAATGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC |
R ARO10 tar | GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATATTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA |
F AGA2 KO tar | TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGTTTATGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC |
R AGA2 KO tar | GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGTTTTGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA |
F AGA1 OE tar | TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAAAAAGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTCCGTTATCAAC |
R AGA1 OE tar | GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGTAAGATCATTTATCTTTCACTGCGGAGAAGTTTCGAACGCCGAAACATGCGCA |
CCasR1-RV | CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTGAACCTCGACTTTCTGGATCAGCC |
CCasR2-FW | AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCTAGCATGGAACCAAGACCCATGACG |
CCasR2-RV | TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTGAACCTCGACTTTCTGGATCAGCC |
CCasR3-RV | TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCTGAACCTCGACTTTCTGGATCAGCC |
CCasR3FW | AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCTAGCATGGAACCAAGACCCATGACG |
CCasR4-FW | GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTAGCATGGAACCAAGACCCATGACG |
CCasR5-FW | GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAGCATGGAACCAAGACCCATGACG |
CCasR5-RV | TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCTGAACCTCGACTTTCTGGATCAGCC |
CCasR1-FW | ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTAGCATGGAACCAAGACCCATGACG |
CCasR4-RV | TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTGAACCTCGACTTTCTGGATCAGCC |
CCasAga1-FW | GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCATGGAACCAAGACCCATGA |
CCasAga1-RV | ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAATGTACTAATTAAAAGATCAGAAAA |
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 |