Table 2.
Cloning/mutagenesis DNA oligos
| Oligo No. | Description | Sequence (5′ → 3′) | Method* |
|---|---|---|---|
| 1 | pcI(T7x-, bicis.) E514TAG F | GCTACCTACCCGAGATAACGC | IPCR |
| 2 | pcI(T7x-, bicis.) E514TAG R: | TAAGCAGCTATCTGGTTCTTAGTG | IPCR |
| 3 | MjTyrRS Y32E F | GATGAAAAATCTGCTGAGATAGGTTTTGAACCAAGTGG | QC |
| 4 | MjTyrRS Y32E R | CCACTTGGTTCAAAACCTATCTCAGCAGATTTTTCATC | QC |
| 5 | MjTyrRS L65H/A67G/H70G F | GATTTAGGCGCCTATTTAAACCAGAAAGG | IPCR |
| 6 | MjTyrRS L65H/A67G/H70G R | ACCCAAATGTATAATTATATCAAATCCAGCATTTTGTAAATC | IPCR |
| 7 | MjTyrRS F108Y/Q109H F | CATCTTGATAAGGATTATACACTGAATGTCTATAGATTG | IPCR |
| 8 | MjTyrRS F108Y/Q109H R | ATATTCACTTCCATAAACATATTTTGCCTTTAACC | IPCR |
| 9 | MjTyrRS D158G/L162G F | TGGTGGCGTTGATGTTGCAGTTG | IPCR |
| 10 | MjTyrRS D158G/L162G R | TAATGAATGGGATTAACCTGCATTATTGGATAGATAACTTCAG | IPCR |
| 11 | MjTyrRS D286R F | GTTTAAAAAATGCTGTAGCTGAAGAACTTATAAAGATTTTAGAG | IPCR |
| 12 | MjTyrRS D286R R | GCATTGGATGCAATTCCTTATTTTTAAATAAACTCTC | IPCR |
| 13 | pcI EcoRI F | ACGTAGCTTGAATTCATGCAGTAGGGAACTGCCAG | |
| 14 | pcI NdeI R: | TTGCTCGTTGCATATGAACCTCCTTAGTACATGCAACC | |
| 15 | CouRS NdeI F | TTCGCTGTGCATATGGACGAATTTGAAATGATAAAGAG | |
| 16 | CouRS EcoRI R | GGCATGAACCGAATTCAGTTATAATCTCTTTCTAATTGGCTCTAAAATC | |
| 17 | pSUP-CouRS BamHI F | ATGCTAAGAGGATCCCGATAAGCTTGGTACCGAGCTC | |
| 18 | pSUP-CouRS SpeI R | TTGGTCTAGCACTAGTTTACAACTTATATCGTATGGGGCTGAC | |
| 19 | pcI(CouRS) BamHI R | TTTACTAGCTGGATCCCAGGAAACAGCTATGACCATGATTAC | |
| 20 | pcI(CouRS) SpeI F | TTACGCATACTAGTGTGCAATGTAACATCAGAGATTTTG | |
| 21 | LIC tRNAopt F: | GAACCACGGAACACAATCAATTCTTGCGGAGAAC | |
| 22 | LIC tRNAopt R | CGACACGGCAAGACCATTCATGTTGTTGCTC | |
| 23 | LIC pSUP tRNA F: | TGGTCTTGCCGTGTCGACGAATTTCTGCCATTCATCC | |
| 24 | LIC pSUP tRNA R | TTGTGTTCCGTGGTTCATCACACTGCTTCCGGTAGTC | |
*IPCR, inverse PCR, QC, QuikChange PCR.