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