TABLE 1.
Synthetic oligonucleotide primers used in this study
| Primera | Sequence (5′-3′) | Significance of underlined sequence |
|---|---|---|
| Sse8387I.f | CCTGCAGGACCAGAATGC | |
| 139.r | GCGGCCGCTCAGTCAGTCTCATTATAATC | NotI site, termination codon |
| 144.r | GCGGCCGCTCAGAATTCTTGGGACCAGTC | NotI site, termination codon |
| 158.r | GCGGCCGCTCATCGGGCAGGGGACACAGA | NotI site, termination codon |
| 163.r | GCGGCCGCTCAATACTCCTCAGCCCATCG | NotI site, termination codon |
| 184.r | GCGGCCGCTCATCGATCAGTGGTGGATGA | NotI site, termination codon |
| 189.r | AACGGTCAATGCGATGTCGACTCAATAGTCATCATACCATCGATC | SalI site, termination codon |
| 223.r | AACGGTCAATGCGATGTCGACTCAAATCTGTCGCACGAATTTCAG | SalI site, termination codon |
| 233.r | AACGGTCAATGCGATGTCGACTCAAGCAGACTCCAGGGACACCTG | SalI site, termination codon |
| 243.r | GCGGCCGCTCACTGTTCTGCCTGAGCTCG | NotI site, termination codon |
| 248.r | AACGGTCAATGCGATGTCGACTCAAAACTCTGCTGCCCACTGTTC | SalI site, termination codon |
| 257.r | AACGGTCAATGCGATGTCGACTCAGGCCTCTGATGTGCCCTGCTG | SalI site, termination codon |
| 262.r | AACGGTCAATGCGATGTCGACTCAGAACTGATCGACCCAGGC | SalI site, termination codon |
| 300.r | AACGGTCAATGCGATGTCGACTCAGGGGTGCGCCTCAGCATCCCG | SalI site, termination codon |
| 305.r | AACGGTCAATGCGATGTCGACTCAATAGTCAGAAAGCCAGGGGTG | SalI site, termination codon |
| 328.r | AACGGTCAATGCGATGTCGACTCAGTGGTCACGCAGGGGGTTCTC | SalI site, termination codon |
| 632.r | AGTCGACTCACTGGGGCAGGCCAAACATAGC | SalI site, termination codon |
| 118.f | TCGAATCGTAGGGATCCATGTGGGCCCAGGAGTTTCTTGCG | BamHI site, initiation codon |
| 123.f | TCGAATCGTAGGGATCCATGCTTGCGGCTGGAGATGCTGTG | EcoRI site, initiation codon |
| 140.f | TCGAATCGTAGGGATCCATGTGGTCCCAAGAATTCATCGTC | BamHI site, initiation codon |
| 145.f | TCGAATCGACTGAATTCATGATCGCTGAAGTCACAGACCCA | EcoRI site, initiation codon |
| 159.f | TCGAATCGACTGAATTCATGTGGGCTGAGGAGTATCTGGAG | EcoRI site, initiation codon |
| 164.f | TCGAATCGACTGAATTCATGCTGGAGCAGTCTGAGGAGAAG | EcoRI site, initiation codon |
| 185.f | TCGAATCGACTGAATTCATGTGGTATGATGACTATCATCCC | EcoRI site, initiation codon |
| 190.f | TCGAATCGACTGAATTCATGCATCCCGAGGAGGATCTGCAG | EcoRI site, initiation codon |
| 206.f | TCGAATCGACTGAATTCATGAAGGTGGACGACCCCAAATTG | EcoRI site, initiation codon |
| 244.f | TCGAATCGACTGAATTCATGTGGGCAGCAGAGTTTATACAG | EcoRI site, initiation codon |
| 249.f | TCGAATCGACTGAATTCATGATACAGCAGCAGGGCACATCA | EcoRI site, initiation codon |
| 258.f | TCGAATCGACTGAATTCATGTGGGTCGATCAGTTCACAAGG | EcoRI site, initiation codon |
| 263.f | TCGAATCGACTGAATTCATGACAAGGTCAGGAAACACGTCT | EcoRI site, initiation codon |
| 306.f | AGAATTCATGGATGACCTCACATCTGCTTCC | EcoRI site, initiation codon |
| Sse8387I.f-2 | TAGGTTCTGAAGATGAGT | |
| W118A/F122A.f | TCTGAGAATGCGGCCCAGGAGGCTCTTGCGGCTGGA | Trp118 and Phe122 mutated to Ala |
| W118A/F122A.r | TCCAGCCGCAAGAGCCTCCTGGGCCGCATTCTCAGA | Trp118 and Phe122 mutated to Ala |
| W140A/F144A.f | GAGACTGACGCGTCCCAAGAAGCCATCGCTGAAGTC | Trp140 and Phe144 mutated to Ala |
| W140A/F144A.r | GACTTCAGCGATGGCTTCTTGGGACGCGTCAGTCTC | Trp140 and Phe144 mutated to Ala |
| W159A/Y163A.f | CCTGCCCGAGCGGCTGAGGAGGCTCTGGAGCAGTCT | Trp159 and Tyr163 mutated to Ala |
| W159A/Y163A.r | AGACTGCTCCAGAGCCTCCTCAGCCGCTCGGGCAGG | Trp159 and Tyr163 mutated to Ala |
| W185A/Y189A.f | ACTGATCGAGCGTCTGATGACGCTCATCCCGAGGAG | Trp185 and Tyr189 mutated to Ala |
| W185A/Y189A.r | CTCCTCGGGATGAGCGTCATCATACGCTCGATCAGT | Trp185 and Tyr189 mutated to Ala |
| Mut-RV-HA | AGTTCAGATAACGCGGCCGCTCACAATGATGCGTAATCTGGTACGTCGTATGGATAGCACTGTTCTGCCTGAGCTCG | NotI site, termination codon, HA epitope |
| KpnI.r | GAGTAGCGCAGCCAGTC | |
| PEX13.f | GCGGATCCTCTAGAGCCATGGCGTCCCAGCCGCCACC | BamHI site |
| PEX13SH3.f | GCGAATTCACTCACAGTGATGAAG | EcoRI site |
| PEX13N.r | ATGCGGCCGCTCACTTTCAATGGACTGAAATGC | NotI site, termination codon |
| M13.r | GTACCAGTATCGACAAAGG | |
| S593W.f | GGCGCCATGTGGGAGAACATCTGGAGT | Ser593 mutated to Trp |
| S593W.r | ACTCCAGATGTTCTCCCACATGGCGCC | Ser593 mutated to Trp |
| 1183.f | AACGAGTCCCTGCAGCG | |
| PEX5stopHA.r | AGTTCAGATAACGCGGCCGCTCACAATGATGCGTAATCTGGTACGTCGTATGGATAGCACTGGGGCAGGCCAAACATAGC | NotI site, termination codon, HA epitope |
| HsCatalase.f | GCGCGTCGACGCTGACAGCCGGGATC | Sal I site |
| HsCatalase.r | GCGCGTCGACTAGTCACAGATTTGCCTTC | SalI site, termination codon |
| 1148.f | GACACTCACCGCCATCGCCTG | |
| ΔKANL.r | GCGCCTCGAGTCACTCCCTTGCCGCCAAGTGAGA | XhoI site, termination codon |
| GFP-SKL.f | CCGACCATGGTGAGCAAGGGC | NcoI site |
| GFP-SKL.r | GGCGTC GACTAGAGCTTGGACTGCAGGTGATGGTGATGGTGATGGAGTCCGGACTTGTA | SalI site, LQSKLTer, His6 |
a
f and r, sense and antisense primers, respectively.