Template:Groningen/Primers
Primers used during this project
| 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 |