Table 2.
Primer sets used to generate pdfr-myc flies
| name | sequence | usage | |
|---|---|---|---|
| 9 | pdfr-LA-AscI-F | AGGCGCGCCGTTTCCTGGCAAGTTTGCGC | subclone the left homologous arm into P[acman] |
| 10 | pdfr-LA-BamHI-R | CGCGGATCCGATGCAATGCATTTGTTACTGC | |
| 11 | pdfr-RA2-BamH1-F | CGCGGATCCGATCGTGATTTGAGAAGAGCTC | subclone the right homologous arm into P[acman] |
| 12 | pdfr-RA2-Pac1-R | ACCTTAATTAACATGATTACGAGAGAGATGGG | |
| 13 | pacman MCS-F | TTTAAACCTCGAGCGGTCCGTTATC | confirm both homologous arm subclones |
| 14 | pacman MCS-R | CTAAAGGGAACAAAAGCTGGGTAC | |
| 15 | pdfr-5′ check-R | GACACTGCGTTGTTGTCATCTAATTG | confirm step (1) gap repair |
| 16 | pdfr-3′ check-F2 | GTCTGTAGTGAAAGAATGCGC | |
| 17 | pdfr-LA-seq-F | GCATGGCCTGATCACTACAG | |
| 18 | pdfr-RA2- seq-R | GATTAACACAGAATGACAGAGG | |
| 19 | 1KB 3′ pdfr F | ATTTGCGGCCGCCGCTTTAATCACCTATTCACCC | subclone Cterm 1 kB into pUASTmyc vector |
| 20 | 1kB 3′ pdfr R | ATTTGCGGCCGCTTCTGCTCTGACAACTCAAATACAA | |
| 21 | pdfr C+myc F | GGCTAGCCGCTTTAATCACCTATTCACC | subclone 1 kB+myc into 3loxPNeoR vector |
| 22 | pdfr C+myc R | CTCTAGAGGCTAGAGAGGCCTTG | |
| 23 | pdfr C 500R F | ACCTTAATTAAACCAAAACCTAGCCTAACTAAT | subclone C term 500 bp the right homolgous arm |
| 24 | pdfr C 500R R | CCTCGAGTGAATGTATTTGCGTGTCCAG | |
| 25 | Neo F | AATATGTATCCGCTCATGAGAC | check step (2) homologous recombination |
| 26 | Neo R | CACCGTGCGTTTTATTCTGTC | |
| 27 | Neo 5′ R | GTCTCATGAGCGGATACATATT | |
| 28 | Neo 3′ F | GACAGAATAAAACGCACGGTG | |
| 29 | myc F | GCGGCCGCGGCTCGACTCCCATCGA | check step (2) homologous recombination and check step (3) removal of NeoR |
| 30 | pdfr LJ | GATCATTCGCCTGACCTTGTA | |
| 31 | pdfr LLB | TACTGTGGTGGTGGGCTACA | |
| 32 | attP-F | CTTCACGTTTTCCCAGGTCAGAAG | check step (4) recombination between attB/attP |
| 33 | attP-R | GTCGCGCTCGCGCGACTGACGGTC | |
| 34 | attB-F | GTCGACGATGTAGGTCACGGTC | |
| 35 | attB-R | TCGACATGCCCGCCGTGACCGTC |