Template:Groningen/Primers
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 |