TABLE 2.
Vectors and primers used in this study
| Vector or primer name | Description | Sequence (5′–3′)a |
Reference or source | |
|---|---|---|---|---|
| Forward | Reverse | |||
| pCMW75 | Ptac-qrgB (pEVS141 backbone) | 23 | ||
| pCMW98 | Ptac-qrgB* (pEVS141 backbone) | 23 | ||
| pCMW121 | Ptac-vc1086 (pEVS141 backbone) | 23 | ||
| pCMW126 | Ptac-vc1086* (pEVS141 backbone) | 23 | ||
| pBRP31 | Promoterless GFP control | This study | ||
| pBRP34 | P2-TS | ATCAGGTACCTATATTTGAAAGCTTGTCAC | ATCACTCGAGCCTGATAAAAATTAATCATTCTTGG | This study |
| pBRP35 | P4-TS; EMSA probe P4 template | ATCAGGTACCACACTTTGTTGACTCATCATTG | ATCACTCGAGCTGTTAGTCTCGGAGTATTG | This study |
| pBRP50 | P3-TS; RNA probe P2 template, EMSA probe P3 template | ATCAGGTACCGCTCATTCTCACATTTGAAATA | ATCACTCGAGCTGTGCGTGACATTTTTCCTG | This study |
| pBRP71 | P2-TS VpsR binding site deletion; RNA probe P1 template, EMSA probe P3 template | ATCAGGTACCTATATTTGAAAGCTTGTCAC | ATCACTCGAGCCCTACTAATAATACTCGCAC | This study |
| pBRP91 | P3-TS VpsR binding site deletion; RNA probe P2 template, EMSA probe P3 template | ATCAGGTACCGTGAGAATGACCCAAGAATG | ATCACTCGAGCTGTGCGTGACATTTTTCCTG | This study |
| pBRP102 | P1-TS; EMSA probe P1 template | ATCAGGTACCGCAGAATACTTCTCTCCACC | ATCACTCGAGCGTTATGAATTGTCAAGAAAGTT | This study |
| pBRP127 | Ptac-tfoY (pEVS141 backbone) | This study | ||
| pBRP175 | Ptac-vpsR qrgB (pEVS141 backbone) | This study | ||
| pBRP179 | Allelic exchange vector, Vc2(G20U-C92U); PCR template for in vitro transcription | ACGCACAGTGCAAACCATTCb | This study | |
| pBRP376 | P1234-TL-FULL gfp reporter fusion | This study | ||
| pBRP379 | pBRP376 with G20U mutation at Vc2 | This study | ||
| pEVS141 | Protein overexpression control | 27 | ||
| pDL1711 | EMSA probe vpsT template | CMW234, 6-FAM–ATTTTGCGGCCGCAACTAGAc | CMW235, 6-FAM–CCGCGGTGGCGGCCGCTCTAc | 8 |
a
For each vector constructed for this study, sequences are given for the primers used in PCR to generate vector inserts. For primer sequences, restriction endonuclease sites are underlined.
b
Mutagenesis sequence.
b
Primer name is given; “forward” and “reverse” do not apply. 6-FAM, 6-carboxyfluorescein.