Primers used to generate mP2X4 constructs carrying pHluorin
| Primer | Sequence (5′ to 3′) |
| P2X4 extreme flanking primers | |
| P2X4 extreme 5′ | AGTTAAGCTTGGTACCGAGCTCGGATCC |
| P2X4 extreme 3′ | TATGCGGCCGCTCACTTGTCATCGTCATCCTTGTAATCCTGGTCCGTCTCTCCGGAAAG |
| P2X4 insertion site primers | |
| P2X4-pHluorin79 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCCTGAGAAGTGTTGGTCACAGCCACACCTTTGGCTTTGG |
| P2X4-pHluorin79 B-5′ | GGAGCTGGAGCTATCGATTATGAATTCGCTGGAGCTGGACTTGGATTCCGGATCTGGGACGTGGCCGACTATGTGG |
| P2X4-pHluorin82 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCGAATCCAAGCTGAGAAGTGTTGGTCAC |
| P2X4-pHluorin82 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGACGGATCTGGGACGTGGCCGACTATG |
| P2X4-pHluorin123 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCCTTATCAGGAATCTCTGGACAGGTACC |
| P2X4-pHluorin123 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGAACCAGCATTTGCGATTCAGACGCCAAC |
| P2X4-pHluorin156 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCAGACGCGTTGAATGGAACACACCTTCC |
| P2X4-pHluorin156 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGAGTGAAGACCTGTGAGGTGGCCGCATGG |
| P2X4-pHluorin207 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCGAGGATATTCCTCTTGCTGAAATTAAAC |
| P2X4-pHluorin207 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGACCCAACATTACCACCTCCTACCTCAAG |
| P2X4-pHluorin222 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCCGCATTATAAATGCACGACTTGAGG |
| P2X4-pHluorin222 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGACGGACGGATCCCTTCTGCCCCATATTCC |
| P2X4-pHluorin224 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCCGTCCGCGCATTATAAATGCACGAC |
| P2X4-pHluorin224 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGAGATCCCTTCTGCCCCATATTCCGTC |
| P2X4-pHluorin304 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCAAGGTCCCTGTAGTACTTGGCAAACC |
| P2X4-pHluorin304 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGAGCTGGCAATGAGCAACGCACACTCACC |
| P2X4-pHluorin307 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCAGACGCGTTGAATGGAACACACCTTCC |
| P2X4-pHluorin307 B-5′ | GAGCTATCGATTATGAATTCGCTGGAGCTGGAGAGCAACGCACACTCACCAAGGCATATG |
| pHluorin amplification primers | |
| pHluorin Cla1 5′ | TATATCGATGGAGGAGCTGGAGGAGCTAGGAGTAAAGGAGAAGAACTTTTCACTGG |
| pHluorin EcoR1 3′ | ATAGAATTCAGCTCCTCCAGCTCCTCCTTTGTATAGTTCATCCATGCCATGTGTAATCC |
| Primer | Sequence (5′ to 3′) |
| P2X4 extreme flanking primers | |
| P2X4 extreme 5′ | AGTTAAGCTTGGTACCGAGCTCGGATCC |
| P2X4 extreme 3′ | TATGCGGCCGCTCACTTGTCATCGTCATCCTTGTAATCCTGGTCCGTCTCTCCGGAAAG |
| P2X4 insertion site primers | |
| P2X4-pHluorin79 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCCTGAGAAGTGTTGGTCACAGCCACACCTTTGGCTTTGG |
| P2X4-pHluorin79 B-5′ | GGAGCTGGAGCTATCGATTATGAATTCGCTGGAGCTGGACTTGGATTCCGGATCTGGGACGTGGCCGACTATGTGG |
| P2X4-pHluorin82 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCGAATCCAAGCTGAGAAGTGTTGGTCAC |
| P2X4-pHluorin82 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGACGGATCTGGGACGTGGCCGACTATG |
| P2X4-pHluorin123 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCCTTATCAGGAATCTCTGGACAGGTACC |
| P2X4-pHluorin123 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGAACCAGCATTTGCGATTCAGACGCCAAC |
| P2X4-pHluorin156 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCAGACGCGTTGAATGGAACACACCTTCC |
| P2X4-pHluorin156 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGAGTGAAGACCTGTGAGGTGGCCGCATGG |
| P2X4-pHluorin207 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCGAGGATATTCCTCTTGCTGAAATTAAAC |
| P2X4-pHluorin207 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGACCCAACATTACCACCTCCTACCTCAAG |
| P2X4-pHluorin222 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCCGCATTATAAATGCACGACTTGAGG |
| P2X4-pHluorin222 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGACGGACGGATCCCTTCTGCCCCATATTCC |
| P2X4-pHluorin224 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCCGTCCGCGCATTATAAATGCACGAC |
| P2X4-pHluorin224 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGAGATCCCTTCTGCCCCATATTCCGTC |
| P2X4-pHluorin304 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCAAGGTCCCTGTAGTACTTGGCAAACC |
| P2X4-pHluorin304 B-5′ | GGAGCTATCGATTATGAATTCGCTGGAGCTGGAGCTGGCAATGAGCAACGCACACTCACC |
| P2X4-pHluorin307 A-3′ | TCCAGCGAATTCATAATCGATAGCTCCAGCTCCAGACGCGTTGAATGGAACACACCTTCC |
| P2X4-pHluorin307 B-5′ | GAGCTATCGATTATGAATTCGCTGGAGCTGGAGAGCAACGCACACTCACCAAGGCATATG |
| pHluorin amplification primers | |
| pHluorin Cla1 5′ | TATATCGATGGAGGAGCTGGAGGAGCTAGGAGTAAAGGAGAAGAACTTTTCACTGG |
| pHluorin EcoR1 3′ | ATAGAATTCAGCTCCTCCAGCTCCTCCTTTGTATAGTTCATCCATGCCATGTGTAATCC |