Table 2.
Primers used during this study
| Construct | Forward | Reverse |
|---|---|---|
| pEC‐K‐3C‐His CAL11–160 | 5′‐ccaggggcccgactcgatggctaacgcggttg‐3′ | 5′‐cagaccgccaccgactgcttattttggcgggctcgc‐3′ |
| CAL11–160‐LL‐841–979 |
5′‐ccaggggcccgactcgatggctaacgcggttg‐3′ 5′‐gcgagcccgccaaaaggcagcagcggcggcagcagcggc ggtggtgacccggattg‐3′ |
5′‐cagaccgccaccgactgcttacttatcaccggag‐3′ 5′‐caatccgggtcaccaccgccgctgctgccgccgctgctgcc ttttggcgggctcgc‐3′ |
| Codon Optimised CAL11–160 W22A/F29A |
5′‐acgaacgttccaaagctgcgtcctctaagatggcgg‐3′ 5′‐ggtcctctaagatggcggatgctgcatccctggaagat‐3′ |
5′‐ccgccatcttagaggacgcagctttggaacgttcgt‐3′ 5′‐atcttccagggatgcagcatccgccatcttagaggacc‐3′ |
| Codon Optimised CAL11–160 W22R/F29R |
5′‐ggtatacgaacgttccaaagctaggtcctctaagatg‐3′ 5′‐ggtcctctaagatggcggatcgtgcatccctggaagat‐3′ |
5′‐catcttagaggacctagctttggaacgttcgtatacc‐3′ 5′‐atcttccagggatgcacgatccgccatcttagaggacc‐3′ |
| Codon Optimised CAL11–160 F43R | 5′‐gcatggaaattgacgtcgcagaacgcgataacctgttccacgg‐3′ | 5′‐ccgtggaacaggttatcgcgttctgcgacgtcaatttccatgc‐3′ |
| pEC‐K‐3C‐His CAL11–50 | 5′‐ccaggggcccgactcgatggctaacgcggttg‐3′ | 5′‐cagaccgccaccgactgcttattcaccgtggaacagg‐3′ |
| 14S CAL1841–979 | 5′‐tacttccaatccaatgcaatgggtggtgacccggattg‐3′ | 5′‐ttatccacttccaatgttatta cttatcaccggagttg‐3′ |
| Codon Optimised CAL1841–979 I900R/K907A/Y908A |
5′‐acagggcctgggcaaaatcaggggcgaacgttg‐3′ 5′aatcatcggcgaacgttgggcgcgtgcggccctgaaataccacattggtagccgttcctt‐3′ |
5′‐caacgttcgcccctgattttgcccaggccctgt‐3′ 5′aaggaacggctaccaatgtggtatttcagggccgcacgcgcccaacgttcgccgatgatt‐3 |
| Non‐Codon Optimised CAL1 W22A/F29A |
5‐gagcgctctaaggccgcgtccagtaaaatggcg‐3′ 5′‐ctggtccagtaaaatggcggatgctgccagcctggaa‐3′ |
5′‐cgccattttactggacgcggccttagagcgctc‐3′ 5′‐ttccaggctggcagcatccgccattttactggaccag‐3′ |
| Non‐Codon Optimised CAL1 W22R/F29R |
5′‐gagcgctctaaggccaggtccagtaaaatgg‐3′ 5′‐ctggtccagtaaaatggcggatcgtgccagcctggaa‐3′ |
5′‐ccattttactggacctggccttagagcgctc‐3′ 5′‐ttccaggctggcacgatccgccattttactggaccag‐3′ |
| Non‐Codon Optimised CAL1 F43R | 5′‐gaaatagatgtggccgagcgcgacaacttgttccacgg‐3′ | 5′‐ccgtggaacaagttgtcgcgctcggccacatctatttc‐3′ |
| Non‐Codon Optimised CAL1 I900R/K907A/Y908A |
5′‐cgagcagggactcggaaagattaggggagaacgttggg‐3′ 5′‐ggagaacgttgggcccgcgcggccctgaagtaccacatcgg‐3′ |
5′‐cccaacgttctcccctaatctttccgagtccctgctcg‐3′ 5′ccgatgtggtacttcagggccgcgcgggcccaacgttctcc‐3′ |
| CENP‐A101–225 S154Q | 5′‐ccaagctgccgttccagcgtctagtgcgcg‐3′ | 5′‐cgcgcactagacgctggaacggcagcttgg‐3′ |
| CENP‐A101–225 M186A | 5′‐caggagtcgtgcgaggcgtacttgacgcagcg ‐3′ | 5′‐cgctgcgtcaagtacgcctcgcacgactcctg ‐3′ |
| CENP‐A101–225 Q190G | 5′‐gagatgtacttgacggggcggctcgccgactc‐3′ | 5′‐gagtcggcgagccgccccgtcaagtacatctc‐3′ |
| CENP‐A101–225 M186A Q190G | 5′‐caggagtcgtgcgaggcgtacttgacggggcg‐3′ | 5′‐cgccccgtcaagtacgcctcgcacgactcctg‐3′ |
| pEC‐K‐3C‐His CENP‐C1264–1411 | 5′‐ccaggggcccgactcgatgggcccggtagtgttc‐3′ | 5′‐cagaccgccaccgactgcttaagaacggatacac‐3′ |