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. 2016 Aug 2;7(6):445–458. doi: 10.1080/21655979.2016.1207019

Table 2.

Oligonucleotides used in this study.

Oligonucleotide Sequence 5′–3′ Primer design Purpose Source
Tps1-F ACTTTCGAAACGATGGGTACTACGGATAACGCTAAGGCGCAACTGACCT BstBI Amplification of tps1 gene This study
Tps1-R ACTGGTACCTCAGTGATGGTGATGGTGATGGTTTTTGGTGGCAGAGGAG KpnI    
VTF TTCGAAACGATGGGTACTAC Positioned from the start codon of the tps1 gene Verification of tps1 gene insertion  
VR AGTGATGGTGATGGTGATGG 6xHis-tag sequence    
L1 CACAAGGTTATCAATATGAAACTTGCGCGATCACCGATTTA Positioned 265 bp from the start codon of the nth1 gene To construct upstream region of nth1 gene  
L2 GTCGACCTGCAGCGTACGTGGACCCTTGTGGTACTGTCAGAGACCTCGTGGACATCCATTCTACTGATTTTTTT 5′ end homologous to 5′ kanMX region    
L3 CGAGCTCGAATTCATCGATGTTGAGCGTGTGTTAATACCCTACGAGACCGACACCGTAGTTGAGATAATATATA 5′ end homologous to 3′ kanMX region To construct downstream region of nth1 gene  
L4 AAGAAAAAGAAACTGTTTTGTCCTTGGAAAAAAAGCACTACCTA Positioned 391 bp from the stop codon of the nth1 gene    
kanMX-F AAAAAAATCAGTAGAATGGATGTCCACGAGGTCTCTGACAGTACCACAAGGGTCCACGTACGCTGCAGGTCGAC 5′ end had 18 bases extensions homologous to the 3′ side of the UP nth1 region. To construct kanMX region Saccharomyces Genome Deletion Project.
kanMX-R TATATATTATCTCAACTACGGTGTCGGTCTCGTAGGGTATTAACACACGCTCAACATCGATGAATTCGAGCTCG 5′ end had 18 bases extensions homologous to the 5′ side of the DOWN nth1 region    
DC-F CCTATCGTTTTCCGTAGAGT Located within the UP nth1 region For amplification of disruption cassette This study
DC-R CCTGAAGTCGAGTAAACAGA Located within the DOWN nth1 region    
A1 CAGGAGCATCGTGTAGGAC Positioned 373 bp from the start codon of the nth1 gene Gene deletion verification primer This study
D1 AAAAGGTCACATGCTTATAATCAAC Positioned 514 bp from the stop codon of the nth1 gene    
B AAACGACGATTACCTTGACTACTTG Located within the coding region of the nth1 gene Gene deletion verification primer Saccharomyces Genome Deletion Project.
kanB CTGCAGCGAGGAGCCGTAAT Positioned 250 bp downstream of the 5 ′-end of the kanMX region    
C TTGTGGTTGAAAAATATGATGTCAC Located within the coding region of the nth1 gene    
kanC TGATTTTGATGACGAGCGTAAT Positioned 587 bp upstream of the 3 ′ end of the kanMX region    
Nth1F ATGAGTCAAGTTAATACAAGCCAAG Positioned from the start codon of the nth1 gene For amplification of nth1 gene This study
Nth1R TAGTCCATAGAGGTTTCTTTCTTG Positioned from the stop codon of the nth1 gene    

The underlined bases encode restriction site and italicized bases encode 6xHis-tag.