Table 2. Primers used in this study.
| Primer | Sequence 5′–3′ |
|---|---|
| pRz S20P FOR | TCTGCCTGCCATGGGCTGTTAATC |
| pRz S20P REV | GATTAACAGCCCATGGCAGGCAGA |
| pRz Q36P | GAGATAACGCCATTACCTACAAAGCCCCGCGCGACAAAAATGCCAGAGAAC |
| pRz A50P | CTGAAGCTGGCGAACGCGCCAATTACTGACATGCAGATGCGTCAGC |
| pRz A62P | CAGATGCGTCAGCGTGATGTTCCTGCGCTCGATGCAAAATACACGAAG |
| pRz T107P FOR | GTGAAGCCACCCCCGCCTCCGGCGTAGATAATG |
| pRz R125P | CTGGCAGACACCGCTGAACCGGATTATTTCACCCTCAGAGAGAGGC |
| pRz E150G | CAACTGGAAGGAACCCAGAAGTATATTTAGGAGCAGTGCAGATAGGGATCC |
| pRE Rz Q151P | CCAGAAGTATATTAATGAGCCGTGCAGATAGGG |
| pRz Q151X | AACAACTGGAAGGAACCCAGAAGTATATTAATGAGTAATGCAGATAGGGATCCGTCGAC |
| pRz C152X | CTGGAAGGAACCCAGAAGTATATTAATGACCAGTAAAGATAGGGATCCGTCGACCTGC |
| pRz R153X | GGAACCCAGAAGTATATTAATGAGCAGTGCTAATAGGGATCCGTCGACCTGCAG |
| pSynRz Linker 101–115 FOR | CGGCAGTGGTAGTGGTAGTGGAAGTCCACGGCTAGCGGAT |
| pSynRz Linker 101–115 REV | CTGCCGCTGCCACTACCGCTTCCTTGGCAAACCGCCTTAATATG |
| SynRz E150R SynRz1 R59E* | CCCTTAGGTACCAGAGAGATTGATGTATGAGCAGAGTCACCGCGATTATCTCCGCTCTGGTTATCTGCATCATCGTCTGCCTGTCATGGGCTGTTAATCATTACCGTGATAACGCCATTACCTACAAAGCCCAGCGCGACAAAAATGCCAGAGAACTGAAGCTGGCGAACGCGGCAATTACTGACATGCAGATGCGTCAGCGTGATGTTGCTGCGCTCGATGCAAAATACACGAAGGAGTTAGCTGACGCCAAGGCGGAGAACGACGCGCTACGGGACGACGTGGCAGCCGGGCGGCGCCGATTACATATTAAGGCGGTTTGCCAATCCGTACGGGAGGCTACTACAGCAAGTGGAGTAGACAACGCGGCAAGTCCACGGCTAGCGGATACTGCCGAGCGAGACTACTTTACACTTAGGGAAAGACTAATCACTATGCAAAAACAACTGGAAGGAACCCAGAAGTATATTAATAGGCAGTGCAGATAGGGATCCAAGGAGTTAGCTGATGCTTAAACTCAAGATGATGCTATGTGTAATGATGTTACCACTTGTTGTAGTTGGGTGTACGAGTAAACAATCGGTATCGCAATGTGTAAAACCGCCCCCACCGCCTGCATGGATCATGCAACCGCCACCTGATTGGCAAACGCCACTAAATGGAATCATATCGCCATCGGAAGAGGGATGAAAGCTTCTGTTTTG |
| pRzART-TMD** | ATATGGTACCAGAGAGATTGATGTATGAGCAGAGTGGTGCTGCTGATTATTGTGGTGGTGGTGGTGGTGGTGGTGATTATTCTGCTGATTATTGTGCATTACCGTGATAACGCCATTACCTACAAAGCCCAGCGCGACAAAAATGCCAGAGAACTGAAGCTGGCGAACGCGGCAATTACTGACATGCAGATGCGTCAGCGTGATGTTGCTGCGCTCGATGCAAAATACACGAAGGAGTTAGCTGACGCCAAGGCGGAGAACGACGCGCTACGGGACGACGTGGCAGCCGGGCGGCGCCGATTACATATTAAGGCGGTTTGCCAATCCGTACGGGAGGCTACTACAGCAAGTGGAGTAGACAACGCGGCAAGTCCACGGCTAGCGGATACTGCCGAGCGAGACTACTTTACACTTAGGGAAAGACTAATCACTATGCAAAAACAACTGGAAGGAACCCAGAAGTATATTAATGAGCAGTGCAGATAGGGATCCGCG |
| pLinkerRz Q100S | GCCGATTACATATTAAGGCGGTTTGCTCAGGAAGCGGTAGTGGCAGC |
| pRz1 25–30 GS-Linker FOR | TCCGGCTCCAAGCCACCACCGCCTCCG |
| pRz1 25–30 GS-Linker REV | GCCGGAGCCCTGCTTTGATGTGCAACCGAC |
| pSynRz1 D45K FOR | AACCGCCACCTAAATGGCAAACGCC |
| pSynRz1 D45K REV | GGCGTTTGCCATTTAGGTGGCGGTT |
| pSynRz1 I54X | GATTGGCAAACGCCACTAAATGGAATCTAATCGCCATCGGAAAGGGGATG |
| pSynRz1 S55X | GGCAAACGCCACTAAATGGAATCATATAGCCATCGGAAAGGGGATGAAAGC |
| pSynRz1 P56X FOR | GGAATCATATCGTGATCGGAAAGGGG |
| pSynRz1 P56X REV | CCCCTTTCCGATCACGATATGATTCC |
| pSynRz1 S57X FOR | CAATCATATCGCCATAAGAAAGGGGATGAAAG |
| pSynRz1 S57X REV | CTTTCATCCCCTTTCTTATGGCGATATGATC |
| pSynRz1 E58X | CCACTAAATGGAATCATATCGCCATCGTAAAGGGGATGAAAGCTTCTGTTTTG |
| pRz1-His P32Q FOR | CGTGAAGCAACCACCGCC |
| pRz1-His P32Q REV | GGCGGTGGTTGCTTCACG |
| pRz1-His P33L FOR | GTGAAGCCACTGCCGCCTCCGGCG |
| pRz1-His P33L REV | CGCCGGAGGCGGCAGTGGCTTCAC |
| pRz1-His P35H FOR | CCACCACCGCATCCGGCGTGG |
| pRz1-His P35H REV | CCACGCCGGATGCGGTGGTGG |
| pRz1-His P36Q FOR | CCACCACCGCCTCAGGCGTGGATAATG |
| pRz1-His P36Q REV | CATTATCCACGCCTGAGGCGCTGGTGG |
| pRz1-His I39V FOR | CCGGCGTGGGTAATGCAGC |
| pRz1-His I39V REV | GCTGCATTACCCACGCCGG |
| pRz1-His P44S FOR | CAGCCTCCCTCCGACTGGC |
| pRz1-His P44S REV | GCCAGTCGGAGGGAGGCTG |
| pRz1-His W46R FOR | CCCCCCGACCGGCAGACAC |
| pRz1-His W46R REV | GTGTCTGCCGGTCGGGGGG |
| pRz1-His L50P FOR | CAGACACCGCCGAACGGGATTATTTC |
| pRz1-His L50P REV | GAAATAATCCCGTTCGGCGGTGTCTG |
| pRz1-His R59E FOR | ATTTCACCCTCAGAGGAAGGCGGCCAC |
| pRz1-His R59 REV | TGGCCGCCTTACTCTGAGGGTGAAATAATCC |
The “*” and “**” symbols indicate a dsDNA gblock (Integrated DNA Technologies) synthesized gene (Genscript) designed with a spanin allele flanked by restriction sites compatible with the pRE plasmid.