TABLE 3.
Oligonucleotides
Oligonucleotide | Sequence (5′-3′) | Use |
---|---|---|
HL446 | atgtgtaggttaagtgatcaaggcgtaatcaggcacatcatatggg taggcgtaatcaggcacatcatatggg tacatcaatacaaactcatcgacaactac |
PCR with HA epitope |
HL211 | tgaaacatttgttttctttgggccggcaaacttattgtttttccaattg | Bottom-strand oligonucleotide creates a unique NgoMI restriction site in gag in pHL1258 |
HL212 | caattggaaaaacaataagtttgccggcccaaagaaaacaaatgtttca | Top strand oligonucleotide creates a unit NgoMI restriction site in gag in pHL1258 |
HL220 | gactacaaggacgacgatgacaag | To generate a 24-bp DNA fragment encoding the FLAG epitope of pHL1276 |
HL221 | cttgtcatcgtcgtccttgtagtc | To generate a 24-bp DNA fragment encoding the FLAG epitope in pHL1276 |
HL199 | gcaactagttcagatcttagtcgaccgatgcataaggatccctgagct | 5′-flanking oligonucleotide of polylinker sequence in library vector pHL1288 |
HL200 | cagggatccttatgcatcggtcgactaagatctgaactagttgctgca | 3′-flanking oligonucleotide of polylinker sequence in library vector pHL1288 |
HL417 | ctgcagcaactagttcagatcttagtcgaccgatgtataaggatccctgagctc | 5′-flanking oligonucleotide in nup124 mutating ATG to a BclI site |
HL340 | gtcgatgagtttgtattgatcactcctgtttcaaaaaatac | 3′-flanking fusion oligonucleotide for overlapping the pair HL417-HL418 with BclI mutation |
HL418 | ctagattcctaacctcgaggtataatcttgcttttgctttggc | 3′-flanking oligonucleotide in nup124 mutating ATG to a BclI site |
HL420 | gtattttttgaaacaggagtgatcaatacaaactcatcgac | 5′-flanking oligonucleotide for overlapping the pair HL417-HL418 with BclI mutation |
HL447 | catcgtaaggatccatttgtcttccaccaatttttgaatt | PCR generation of the 3′ flank of S. pombe his3 gene with pHL1570 as template |
HL440 | gtatatacggccgtctatgcaaagctaacgaatct | PCR generation of the 5′ flank of S. pombe his3 gene with pHL1570 as template |
HL495 | tttggcagatctctttcaacgttt | PCR oligonucleotide 5′ flank for generation of Gag in GST vector |
HL496 | gctttaatcagaggatccatgaaaaactcatcacagaaaagaatt | PCR oligonucleotide 3′ flank for generation of Gag in GST vector |
HL520 | agttagcacaaggataccctcgagtcaataacgtcttttcttgtattttgta | PCR oligonucleotide 5′ flank for generation of an N-terminal portion of Nup124p in GST vector |
HL510× | catgctttaatcagaggatccagaattattcgcgttagtaatcggagccat | PCR oligonucleotide 3′ flank for generation of an N-terminal portion of Nup124p in GST vector |
HL521 | agttagcacaaggataccgtcgactcacaaacgattagtttgcacatcagc | PCR oligonucleotide 5′ flank for generation of C-terminal section of Nup124p in GST vector |
HL501 | catgctttaatcagaggatccaaggagaatgagcccaagccaacg | PCR oligonucleotide 3′ flank for generation of C-terminal section of Nup124p in GST vector |
HL339 | gaattcatgcctcctgtttcaaaaaatac | 5′ flank for 779–1073 nucleotide synthesis of nup124 segment 1 to generate LexA fusions |
HL340 | ctcgaggtataacttgcttttgctttgg | 3′ flank for 779–1073 nucleotide synthesis of nup124 segment 1 to generate LexA fusions |
HL341 | gaattcagaattattcgcgttagtaatcgg | 5′ flank for 1073–1718 nucleotide synthesis of nup124 segment 2 to generate LexA fusions |
HL342 | ctcgagtcatgcattgactaatacaggcttc | 3′ flank for 1073–1718 nucleotide synthesis of nup124 segment 2 to generate LexA fusions |
HL343 | gaattcgaagttcagacagattccaacc | 5′ flank for 1718–2363 nucleotide synthesis of nup124 segment 3 to generate LexA fusions |
HL344 | ctcgagtcacaaacgattagtttgcacaatca | 3′ flank for 1718–2363 nucleotide synthesis of nup124 segment 3 to generate LexA fusions |
HL345 | gaattcaaggagaatgagcccaagcc | 5′ flank for 2363–3008 nucleotide synthesis of nup124 segment 4 to generate LexA fusions |
HL346 | ctcgagtcacttggtaaaattaaatgaaaactg | 3′ flank for 2363–3008 nucleotide synthesis of nup124 segment 4 to generate LexA fusions |
HL347 | gaattcccaaatacggatgccaaaacc | 5′ flank for 3008–3653 nucleotide synthesis of nup124 segment 5 to generate LexA fusions |
HL348 | ctcgagtcaggtagtctttgaagttgcagac | 3′ flank for 3008–3653 nucleotide synthesis of nup124 segment 5 to generate LexA fusions |
HL349 | gaattctctgaaggaaccgctccagc | 5′ flank for 3653–4306 nucleotide synthesis of nup124 segment 6 to generate LexA fusions |
HL350 | ctcgagttaacgttttcttcgacttcgg | 3′ flank 3653–4306 nucleotide synthesis of nup124 segment 6 to generate LexA fusions |
HL414 | gaattcatgaaaaaactcatcacagaaaaggattcgaatggaaatgg | 5′ flank for Gag to generate B42 activation domain fusions |
HL355 | ctcgagtcaataacgtcttttcttgtattttg | 3′ flank for Gag to generate B42 activation domain fusions |
HL613 | ggtattcaattcaatttgggctcttcta actcacaaacaaatgcgcccccgggccg taaaattgctgtgccccgaagtcgaa gaaaacgttccggaggaggtggaggagagctcatgggtattcctaac |
5′-flanking PCR oligonucleotide for generating nup124::pk1(3) GFP SV40 poly(A) ura4+ integration tag |
HL614 | taaaagcgtccgtatgtgtaaacaaaaatgtaataattatt ccttagcaattaatttaaaggacaaagtctatctaaac tatcatatgagaaagtctttgctgatatgcctt |
3′-flanking PCR oligonucleotide for generating nup124::pk1(3) GFP SV40 poly(A) ura4+ integration tag |