TABLE 3.
Primers used in this study
| Primer description | Primer sequence (5′–3′) |
|---|---|
| Fw primer to screen iKO integration (primer 2, Fig. S1) | CCGGAATTCAAGAAAAAATGCCAACGAGTAGTTTTC |
| Fw primer to screen iKO integration (primer 3, Fig. S1) | TGGCCAGGGCGAATTGGGTACCGAGC |
| Fw primer for KI-AMA1-Myc-BirA | CGGGGTACCCAAGCCTGTAAAAGACAGAAGACGTCCTG |
| Rev primer for KI-AMA1-Myc-BirA | CGGCCTGCAGGGTAATCCCCCTCGACCATAACATGTG |
| Fw primer for GST-Rab11A | CGGggatccGAACAAAAACTCATCTCAGAAGAGGATCTGATGGCGGCTAAAGATGAATACTACG |
| Rev primer for GST-Rab11A | gcggccgcTCAGGCGGAACAGCAGCCAC |
| Fw primer annealing downstream of ATG of HXGPRT resistance cassette (primer 1, Fig. S3) | CGACAACACCTTCTACAACGCTG |
| Universal reverse primer for CRISPR/Cas9 guide | AACTTGACATCCCCATTTAC |
| Fw primer for KI-HOOK-HA-DHFR | tacttccaatccaatttagcGCAAAGATGACATGGCGAAGCAGATGATG |
| Rev primer for KI-HOOK-HA-DHFR | tcctccacttccaattttagcCGCCTCCCGAGGTGTGACAGAATC |
| Fw primer for KI-HOOK-3Ty-DHFR | GCGGGTACCGATGGAGGCTTTGAGCAAACACG |
| Rev primer for KI-HOOK-3Ty-DHFR | GCGATGCATGCGCCTCCCGAGGTGTGACAG |
| gRNA for iKO tet system, pSAG1::CAS9-GFP-U6::sgHOOK2 | GAGTCCGCACCTCTACACGGGGTTTTAGAGCTAGAAATAGC |
| Fw primer for iKD Tet system | CTTCGTCAATAATGTTTCATCCATTTCGTTCATGTTTGCGGATCCGGGG |
| Rev primer for iKD Tet system | GTCGAGCAAGGGCGCGTGAGTGAAAGACATCAGGTCCTCCTCGGAGATGA |
| Fw primer to screen iKO integration (primer 1, Fig. S1) | GCGCCATGGTAACCGTTTAGCTGCAACAATCC |
| Rev primer to screen iKO integration (primer 4, Fig. S1) | GCGACTAGTAACTCGAGACCCATTTGAGAACC |
| gRNA for HOOK KO, pSAG1::CAS9-GFP-U6::sgHOOK1 | GACCGAAATGGCGGCCCTGAGTTTTAGAGCTAGAAATAGC |
| Fw primer to screen HOOK-KO | CGTCCCAGACTCAAAGAAGAAATG |
| Rev primer to screen KO | GGTTCCAAGGCGAGTGCAGC |
| Fw primer for KI-FTS-cMyc-HXGPRT and KI-FTS-HA-DHFR | TACTTCCAATCCAATTTAGCATTCCGTCATGGGACAATCTTCGAG |
| Rev primer for KI-FTS-cMyc-HXGPRT and KI-FTS-HA-DHFR | TCCTCCACTTCCAATTTTAGCTTCGGCGTTGAAGAGGTTGGCGCC |
| gRNA for KI-APR1-2Ty, pSAG1::CAS9-GFP-U6::sgAPR1 | ATTCACATCGGCCTATATACGTTTTAGAGCTAGAAATAGC |
| Fw primer for C terminus KI of APR1-Ty in KI-FTS-cMyc and KI-HOOK-HA | GGGACCCCTCCGCCGTGGAGAGTTAAAAGCGCTAGCAAGGGCTC GGG |
| Rev primer for C terminus KI of APR1-Ty in KI-FTS-cMyc and KI-HOOK-HA | CAAAAACTGATACCGAGTGTCGCACTGGCAATACGACTCACTATA GGG |
| gRNA for FTS-mAID-HA and KI-2Ty, pSAG1::CAS9-GFP-U6::sgFTS | GTGTAGCTTGCTGGTGGCCATGTTTTAGAGCTAGAAATAGC |
| Fw Primer, KOD PCR for FTS-mAID and KI-2Ty | AGTACTGGCGCCAACCTCTTCAACGCCGAAGCTAGCAAGGGCTCGGG |
| Rev primer, KOD PCR for FTS-mAID and KI-2Ty | AGAAAAGACTACCAGTAGAACCCGCAAAGCATACGACTCACTATAGGG |
| Rev primer screen integration FTS (primer 2, Fig. S3) | CGCACTCCCGGTACTATTTCC |
| gRNA for HIP-mAID-HA and KI-2Ty, pSAG1::CAS9-GFP-U6::sgHIP | GCCTCGGCGATGCCCCTTTGCGTTTTAGAGCTAGAAATAGC |
| Fw primer, KOD PCR for HIP-mAID and KI-2Ty | CCGTCCGCTTCGTCTTCAACAGCGTCTACGGCTAGCAAGGGCTCGGG |
| Rev primer, KOD PCR for HIP-mAID and KI-2Ty | ATTGACTGCAGGGCTGTAGAGAGCTACGCCATACGACTCACTATAGGG |