Table 2.
Oligonucleotides used to generate pYopH FLAG mCherry.
| Number | Sequence | Dir | Function | Enzyme |
|---|---|---|---|---|
| SMP425 | GGAggatccGCTGCGCGATGTACTGACCCG | >> | pyopH 5′ | BamHI |
| SMP426 | CTCCTCGCCCTTGCTCACCATCTTGTCATCGTCGTCCTTGTAATCCATATGTCCCTCCTTAATTAAATACACGCCTATAC | < < | pyopH-FLAG-mCherry/EGFP (for SOE, 3′, use with SMP425) | |
| SMP427 | GTATAGGCGTGTATTTAATTAAGGAGGGACATatggattacaaggacgacgatgacaagAtggtgagcaagggcgaggag | >> | pyopH-FLAG-mCherry/EGFP (for SOE, 5′, use with SMP428) | |
| SMP428 | GAAgaattcTTACTTGTACAGCTCGTCCATGCCG | < < | mCherry/EGFP 3′ | EcoRI |
| SMP431 | GAAgaattcTTACGCTGCTAACGCGTAATTCTCATCATTCGCTGCCTTGTACAGCTCGTCCATGCCGC | < < | mCherry/EGFP 3′ with ssrA (LAA) tag (use with 425,427) | EcoRI |
| SMP435 | CAGCGAATGATGAGAATTACGCGTTAGTAGCGTAAGAATTCTGTTTCCTGTGTG | >> | LAA->LVA mut. for pMMB67EH-PyopH-FLAG-mCherry (top) | |
| SMP436 | CACACAGGAAACAGAATTCTTACGCTACTAACGCGTAATTCTCATCATTCGCTG | < < | LAA->LVA mut. for pMMB67EH-pyopH-FLAG-mCherry (bottom) | |
| SMP437 | GCAGCGAATGATGAGAATTACGCGgCAGCAGTGTAAGAATTCTGTTTCCTGTGTGAAATTG | >> | LAA->AAV mut. for pMMB67EH-pyopH-FLAG-mCherry (top) | |
| SMP438 | CAATTTCACACAGGAAACAGAATTCTTACACTGCTGCCGCGTAATTCTCATCATTCGCTGC | < < | LAA->AAV mut. for pMMB67EH-PyopH-FLAG-mCherry (bottom) | |
| SMP455 | CAAGGCAGCGAATGATGAGAATTACGCGGCAtCAGtGTAAGAATTCTGTTTCCTGTGTGAAATTGTTATC | >> | LAA->ASV mut. for pMMB67EH-pyopH-FLAG-mCherry (top) | |
| SMP456 | GATAACAATTTCACACAGGAAACAGAATTCTTACACTGATGCCGCGTAATTCTCATCATTCGCTGCCTTG | < < | LAA->ASV mut. for pMMB67EH-pyopH-FLAG-mCherry (bottom) |