Oligonucleotide primers used for the construction of the cysteine mutants of SakSTAR
| Variant . | Primer . | Code . | Sequence . |
|---|---|---|---|
| External | Sense | 818A | 5′-CAG GAA ACA GAA TTC AGG AG-3′ |
| Antisense | 818D | 5′-CAA AAC AGC CAA GCT TCA TTC ATT CAG C-3′ | |
| Antisense | 819D | 5′-CAG GCT GAA AAT CTT CTC TCA TCC GCC | |
| SakSTAR(K96C) | Sense | LJ452 | 5′-TAT GAT AAG AAT |
| Antisense | LJ453 | 5′-TTC TTC TTT TTT | |
| SakSTAR(K102C) | Sense | LJ454 | 5′-AAA AAG AAG AAA |
| Antisense | LJ455 | 5′-TAG GGA AAG | |
| SakSTAR(K109C) | Sense | LJ456 | 5′-CCC TAT AAC AGA |
| Antisense | LJ457 | 5′-GGG ACA ACA AAA | |
| SakSTAR(K135C) | Sense | LJ337 | 5′-GAA ACA GAA TTC AGG AGG CCT CAT ATG TCG AGC TCA TTC GAC AAA GG-3′ |
| SakSTAR(K135C), SakSTAR(K96C, K135C) | Antisense | LJ522 | 5′-CAG CCA AGC TTC ATT CAT TCA GCA TGC TGC AGT TAT |
| SakSTAR(S3C) | Sense | LY459 | 5′-CCT CAT ATG TCA AGT |
| SakSTAR(S2C, S3C) | Sense | LY461 | 5′-CCT CAT ATG TCA |
| Variant . | Primer . | Code . | Sequence . |
|---|---|---|---|
| External | Sense | 818A | 5′-CAG GAA ACA GAA TTC AGG AG-3′ |
| Antisense | 818D | 5′-CAA AAC AGC CAA GCT TCA TTC ATT CAG C-3′ | |
| Antisense | 819D | 5′-CAG GCT GAA AAT CTT CTC TCA TCC GCC | |
| SakSTAR(K96C) | Sense | LJ452 | 5′-TAT GAT AAG AAT |
| Antisense | LJ453 | 5′-TTC TTC TTT TTT | |
| SakSTAR(K102C) | Sense | LJ454 | 5′-AAA AAG AAG AAA |
| Antisense | LJ455 | 5′-TAG GGA AAG | |
| SakSTAR(K109C) | Sense | LJ456 | 5′-CCC TAT AAC AGA |
| Antisense | LJ457 | 5′-GGG ACA ACA AAA | |
| SakSTAR(K135C) | Sense | LJ337 | 5′-GAA ACA GAA TTC AGG AGG CCT CAT ATG TCG AGC TCA TTC GAC AAA GG-3′ |
| SakSTAR(K135C), SakSTAR(K96C, K135C) | Antisense | LJ522 | 5′-CAG CCA AGC TTC ATT CAT TCA GCA TGC TGC AGT TAT |
| SakSTAR(S3C) | Sense | LY459 | 5′-CCT CAT ATG TCA AGT |
| SakSTAR(S2C, S3C) | Sense | LY461 | 5′-CCT CAT ATG TCA |
Numbering and orientation are relative to the mature staphylokinase polypeptide.