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+ | <h5>Synthetic DNA fragments</h5> | ||
<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> | <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><p> </p> | <div class="clear"></div><p> </p> | ||
<br> | <br> | ||
+ | <h5><i>Saccharomyces cerevisiae</i> strains</h5> | ||
<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> | <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><p> </p> | <div class="clear"></div><p> </p> | ||
<br> | <br> | ||
+ | <h5>Plasmids</h5> | ||
<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> | ||
Revision as of 21:26, 17 October 2018
Primers, plasmids, and strains used during this project
Primers
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 fragments
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" |
Saccharomyces cerevisiae strains
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
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 |