Template:Groningen/Primers

Sequences

CRISPR-Cas Primerssequence
588 FW CTTGAAAACTTACGCTCATCTCTTCGACG
589 RVCGGACTTAAGAAAATCCAGGATTGTCTTTCC
6006 FW GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC
6005 RVGATCATTTATCTTTCACTGCGGAGAAG
F ARO10 tar TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAACAATGGTTTTAGAGCTAGAAATA
GCAAGTTAAAATAAGGCTAGTCCGTTATCAAC
R ARO10 tar GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATATTGATCATTTATCTTTCACTGC
GGAGAAGTTTCGAACGCCGAAACATGCGCA
F AGA2 KO tar TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGTTTATGTTTTAGAGCTAGAAATAGCAA
GTTAAAATAAGGCTAGTCCGTTATCAAC
R AGA2 KO tar GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGTTTTGATCATTTATCTTTCACTGCGGAG
AAGTTTCGAACGCCGAAACATGCGCA
F AGA1 OE tar TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAAAAAGTTTTAGAGCTAGAAATAGCAAGTTA
AAATAAGGCTAGTCCGTTATCAAC
R AGA1 OE tar GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGTAAGATCATTTATCTTTCACTGCGGAGAAGT
TTCGAACGCCGAAACATGCGCA
CCasR1-RV CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTGAAC
CTCGACTTTCTGGATCAGCC
CCasR2-FW AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCTAGCAT
GGAACCAAGACCCATGACG
CCasR2-RV TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTGAAC
CTCGACTTTCTGGATCAGCC
CCasR3-RV TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCTGAACC
TCGACTTTCTGGATCAGCC
CCasR3FW AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCTAGCATG
GAACCAAGACCCATGACG
CCasR4-FW GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTAGCATGG
AACCAAGACCCATGACG
CCasR5-FW GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAGCATGG
AACCAAGACCCATGACG
CCasR5-RV TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCTGAACCTC
GACTTTCTGGATCAGCC
CCasR1-FW ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTAGCATG
GAACCAAGACCCATGACG
CCasR4-RV TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTGAAC
CTCGACTTTCTGGATCAGCC
CCasAga1-FW GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCA
TGGAACCAAGACCCATGA
CCasAga1-RV ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAATGTAC
TAATTAAAAGATCAGAAAA
Cellulosome primerssequence
BGLI-fragA-fw TAGTAATGCAGACACTTGCG
BGLI-fragA-rvCAACAGCGGTCTACTGAATC
BGLI-fragB-fwCACTAGCTCAGATTCAGTAGACC
BGLI-fragB-rvCGAGATGGACCGCAAG
BGLI1-FWTTTGGATCCGTCTAGATGAA
BGL1-RVAAACTCGAGTACTAGTATTC
BGL1A-FWTTTGGATCCGTCTAGATGAGATTTCCTTCAATTTTTAC
BGL1A-RVAAACTCGAGTACTAGTACTAATAAGGTAGGTGGGGTA
CBHII1-FWTTTGGATCCGTCTAGATGAA
CBHII1-RVAAACTCGAGTACTAGTAACT
CIPA3A-FWTTTGGATCCGTCTAGATGCA
CIPA3A-RVAAAGTCGACCTCGAGTACTA
CIPA3B-FWTTTGGATCCGTCTAGATGCAGTTACTTCGCTGTTT
CIPA3B-RVAAACTCGAGTACTAGTATTAGTGATGGTGATGGTGATGTTCGAAGGGCCCATTCGAAT
CIPA3C-RVAAAGTCGACCTCGAGTATCA
EGII1-FWTTTGGATCCGTCTAGATGAA
EGII1-RVAAACTCGAGTACTAGTAGAC
Bio-His-EG-FW1TATTTGAATTCGCGGCCGCTTCTAGATGCATCATCACCATCACCATG
Bio-His-EG-RV1TATTTCTGCAGCGGCCGCTACTAGTATTACTTTCTTGCGAGACACGAG
Bio-ss1-HEGII-FW1TATTTGAATTCGCGGCCGCTTCTAGATGAAGGTTTTGATTGTCTTG
Int-ss1-HEGII-FW1TTTGGATCCGTCTAGATGATGAAGGTTTTGATTGTCTTG
Int-ss1-HEGII-RV1AAACTCGAGTACTAGTATTACTTTCTTGCGAGACACGAGC

Sequences

CRISPR-Cas Primerssequence
588 FW CTTGAAAACTTACGCTCATCTCTTCGACG
589 RVCGGACTTAAGAAAATCCAGGATTGTCTTTCC
6006 FW GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC
6005 RVGATCATTTATCTTTCACTGCGGAGAAG
F ARO10 tar TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAATATACGAACGAAACAATGGTTTTAGAGCTAGAAATA
GCAAGTTAAAATAAGGCTAGTCCGTTATCAAC
R ARO10 tar GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACCATTGTTTCGTTCGTATATTGATCATTTATCTTTCACTGC
GGAGAAGTTTCGAACGCCGAAACATGCGCA
F AGA2 KO tar TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCAAAACATACTGTGTGTTTATGTTTTAGAGCTAGAAATAGCAA
GTTAAAATAAGGCTAGTCCGTTATCAAC
R AGA2 KO tar GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACATAAACACACAGTATGTTTTGATCATTTATCTTTCACTGCGGAG
AAGTTTCGAACGCCGAAACATGCGCA
F AGA1 OE tar TGCGCATGTTTCGGCGTTCGAAACTTCTCCGCAGTGAAAGATAAATGATCTTACGTTCATCTTTGAAAAAGTTTTAGAGCTAGAAATAGCAAGTTA
AAATAAGGCTAGTCCGTTATCAAC
R AGA1 OE tar GTTGATAACGGACTAGCCTTATTTTAACTTGCTATTTCTAGCTCTAAAACTTTTTCAAAGATGAACGTAAGATCATTTATCTTTCACTGCGGAGAAGT
TTCGAACGCCGAAACATGCGCA
CCasR1-RV CGTGTAATCTGTGACAATTTCGATGCGTACACTGCGGCCCCGAAGTAGCATAGCTTATCTGGATCTGAAC
CTCGACTTTCTGGATCAGCC
CCasR2-FW AGATAAGCTATGCTACTTCGGGGCCGCAGTGTACGCATCGAAATTGTCACAGATTACACGCGCTCTAGCAT
GGAACCAAGACCCATGACG
CCasR2-RV TGAAATGTTATTATGTACCTCAGTACGCGCTAACATCCCTGGGTTACGCATCACTACATTGGATCTGAAC
CTCGACTTTCTGGATCAGCC
CCasR3-RV TGTATCGAGCGATGTACGAATACTCTTGCCGAGTCATAGCAGTGCAGTAGAGCCTGACCCGGATCTGAACC
TCGACTTTCTGGATCAGCC
CCasR3FW AATGTAGTGATGCGTAACCCAGGGATGTTAGCGCGTACTGAGGTACATAATAACATTTCACGCTCTAGCATG
GAACCAAGACCCATGACG
CCasR4-FW GGGTCAGGCTCTACTGCACTGCTATGACTCGGCAAGAGTATTCGTACATCGCTCGATACACGCTCTAGCATGG
AACCAAGACCCATGACG
CCasR5-FW GCTCTCAACAGTCACTTGTGTGCCAAAATATTATACCTACTTCATACATTTTCAATTAAGCGCTCTAGCATGG
AACCAAGACCCATGACG
CCasR5-RV TAATCTTACACTAGTTTATAAGTGTCACGAACGATGAAATGGAAACTTGGGGGGGAAGGCGGATCTGAACCTC
GACTTTCTGGATCAGCC
CCasR1-FW ACAAGTTGACGCGACTTCTGTAAAGTTTATTTACAAGATAACAAAGAAACTCCCTTAAGCCGCTCTAGCATG
GAACCAAGACCCATGACG
CCasR4-RV TGTTAATGAACAGAAAACGAACAATTGGTAGCAGTGTTTTATAATTGCGCCCACAAGTTTGGATCTGAAC
CTCGACTTTCTGGATCAGCC
CCasAga1-FW GCGCAGAATAATGATCTATCATCAATAATTATCGTGAACTTTTCCAGTATTTACGTTCATGCA
TGGAACCAAGACCCATGA
CCasAga1-RV ATTAGTTAATCCCAACAATATTGTGAACAGGTAGGTAAAATGAGCGAAAGATAATGTCATACGAATGTAC
TAATTAAAAGATCAGAAAA

Cellulosome primerssequence
BGLI-fragA-fw TAGTAATGCAGACACTTGCG
BGLI-fragA-rvCAACAGCGGTCTACTGAATC
BGLI-fragB-fwCACTAGCTCAGATTCAGTAGACC
BGLI-fragB-rvCGAGATGGACCGCAAG
BGLI1-FWTTTGGATCCGTCTAGATGAA
BGL1-RVAAACTCGAGTACTAGTATTC
BGL1A-FWTTTGGATCCGTCTAGATGAGATTTCCTTCAATTTTTAC
BGL1A-RVAAACTCGAGTACTAGTACTAATAAGGTAGGTGGGGTA
CBHII1-FWTTTGGATCCGTCTAGATGAA
CBHII1-RVAAACTCGAGTACTAGTAACT
CIPA3A-FWTTTGGATCCGTCTAGATGCA
CIPA3A-RVAAAGTCGACCTCGAGTACTA
CIPA3B-FWTTTGGATCCGTCTAGATGCAGTTACTTCGCTGTTT
CIPA3B-RVAAACTCGAGTACTAGTATTAGTGATGGTGATGGTGATGTTCGAAGGGCCCATTCGAAT
CIPA3C-RVAAAGTCGACCTCGAGTATCA
EGII1-FWTTTGGATCCGTCTAGATGAA
EGII1-RVAAACTCGAGTACTAGTAGAC
Bio-His-EG-FW1TATTTGAATTCGCGGCCGCTTCTAGATGCATCATCACCATCACCATG
Bio-His-EG-RV1TATTTCTGCAGCGGCCGCTACTAGTATTACTTTCTTGCGAGACACGAG
Bio-ss1-HEGII-FW1TATTTGAATTCGCGGCCGCTTCTAGATGAAGGTTTTGATTGTCTTG
Int-ss1-HEGII-FW1TTTGGATCCGTCTAGATGATGAAGGTTTTGATTGTCTTG
Int-ss1-HEGII-RV1AAACTCGAGTACTAGTATTACTTTCTTGCGAGACACGAGC

Sequencing primerssequence
genseqFWGCAGACGACTCGAATAAG
genseqRVAGCTCGTCAATAGTGGAGATTAC
VR primer fw ATTACCGCCTTTGAGTGAGC
VR primer rvTGCCACCTGACGTCTAAGAA
M13 fwGTAAAACGACGGCCAGT
M13 rvCAGGAAACAGCTATGAC
EGII-seq-fwGGCTGTACCACAGATGG
EGII-seq-rvGTTCTGTCGGAGCCAAG
BGL1-seq-paper-FWGCAACAGCCAAGCATTAC
BGL1-seq2-paper-FW ACCACAGCGAGGTTATTC
PAL2-seq-ver-RVGTGGTGCTCATCAGTTTC
PAL2-seq-fw-internalCTGCGTGGCACCATTAC

Zeocin cassette primerssequence
Zcas1-FWTATTTGAATTCGCGGCCGCTTCTAGAGTCCAGAAAGTCGAGGTTCAG
Zcas2-FW TATTTGAATTCGCGGCCGCTTCTAGAGGGGCTGATCCAGAAAGTCGA
Zcas3-FWTATTTGAATTCGCGGCCGCTTCTAGAGCCAGAAAGTCGAGGTTCAG
Zcas4-FW TATTTGAATTCGCGGCCGCTTCTAGAGGATCCAGAAAGTCGAGGTTCAG
Zcas1-RVTATTTCTGCAGCGGCCGCTACTAGTATTATGCTTCCGGCTCGTATG
Zcas2-RV TATTTCTGCAGCGGCCGCTACTAGTACTTCCGGCTCGTATGTTGTG
Zcas3-RVTATTTCTGCAGCGGCCGCTACTAGTAGCTCGGAATTAACCCTCAC

Synthetic DNA sequencesSource
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"

StrainsSource
BJ1991"Molecular Cell Biology group, RUG"
BY4741"Molmic group, RUG"
BJC3CBThis work
BJC1CBThis work
YPH499"Molecular Cell Biology group, RUG"
YPHPAL2This work
YPHCONThis work

PlasmidsSource
pYD1-CipA3-EGIIFrom Wen et al. 2010
pYD1-CipA1-EGIIFrom Wen et al. 2010
pRS425-CBHII-BGL1From Wen et al. 2010
p414 KanMX TEF1-Cas9-Tcyc"Molmic group, RUG"
pMEL16 OR1"Molmic group, RUG"
pMEL16-ARO10tarThis work
pMEL16-AGA2tarThis work
pMEL16-AGA1tarThis work
pUC57-PAL2"Roelfes group, RUG"
pRS313+ s promHxt7-MCS-terHxt7"Molmic group, RUG"
pRS313-Hxt7-PAL2This work


Sequencing primerssequence
genseqFWGCAGACGACTCGAATAAG
genseqRVAGCTCGTCAATAGTGGAGATTAC
VR primer fw ATTACCGCCTTTGAGTGAGC
VR primer rvTGCCACCTGACGTCTAAGAA
M13 fwGTAAAACGACGGCCAGT
M13 rvCAGGAAACAGCTATGAC
EGII-seq-fwGGCTGTACCACAGATGG
EGII-seq-rvGTTCTGTCGGAGCCAAG
BGL1-seq-paper-FWGCAACAGCCAAGCATTAC
BGL1-seq2-paper-FW ACCACAGCGAGGTTATTC
PAL2-seq-ver-RVGTGGTGCTCATCAGTTTC
PAL2-seq-fw-internalCTGCGTGGCACCATTAC
Zeocin cassette primerssequence
Zcas1-FWTATTTGAATTCGCGGCCGCTTCTAGAGTCCAGAAAGTCGAGGTTCAG
Zcas2-FW TATTTGAATTCGCGGCCGCTTCTAGAGGGGCTGATCCAGAAAGTCGA
Zcas3-FWTATTTGAATTCGCGGCCGCTTCTAGAGCCAGAAAGTCGAGGTTCAG
Zcas4-FW TATTTGAATTCGCGGCCGCTTCTAGAGGATCCAGAAAGTCGAGGTTCAG
Zcas1-RVTATTTCTGCAGCGGCCGCTACTAGTATTATGCTTCCGGCTCGTATG
Zcas2-RV TATTTCTGCAGCGGCCGCTACTAGTACTTCCGGCTCGTATGTTGTG
Zcas3-RVTATTTCTGCAGCGGCCGCTACTAGTAGCTCGGAATTAACCCTCAC
Synthetic DNA sequencesSource
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"
StrainsSource
BJ1991"Molecular Cell Biology group, RUG"
BY4741"Molmic group, RUG"
BJC3CBThis work
BJC1CBThis work
YPH499"Molecular Cell Biology group, RUG"
YPHPAL2This work
YPHCONThis work
PlasmidsSource
pYD1-CipA3-EGIIFrom Wen et al. 2010
pYD1-CipA1-EGIIFrom Wen et al. 2010
pRS425-CBHII-BGL1From Wen et al. 2010
p414 KanMX TEF1-Cas9-Tcyc"Molmic group, RUG"
pMEL16 OR1"Molmic group, RUG"
pMEL16-ARO10tarThis work
pMEL16-AGA2tarThis work
pMEL16-AGA1tarThis work
pUC57-PAL2"Roelfes group, RUG"
pRS313+ s promHxt7-MCS-terHxt7"Molmic group, RUG"
pRS313-Hxt7-PAL2This work
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