TABLE 3.
Primers used in this study
| Primer | Sequence | Purpose |
|---|---|---|
| dest-FWD | TAGGGATAACAGGGTAATCAGTCGTGTCATCTGATTACCTGGCGGAAATTAAACTAAGAGAGAGCTCTAAATCATGAAAAATTTATTTGCTTTGTG | Construction of pET30a-dest |
| dest-REV | CGAATTAGCCTGCCAGCCCTGTTTTTATTAGTGCATTTTGCGCGAGGTCAAAATAAATTTTTCATGATTT | Construction of pET30a-dest |
| I-SceI-LP-T5-FWD | TCCGATTAGGGATAACAGGGTAATGTGTAGGCTGGAGCTGCTTCTACGGCCCCAAGGTCCAAACGGTGAAAATCATGAAAAATTTATTTGCTTTGTG | Construction of pET30a-LP-pcaIJFK, pET30a-LP-control |
| LP-IJFK-REV | ACAAGGCCATGGCTGATCATATGAATATCCTCCTTAGTTGGCTTCAGGGATGAGGCGCCATCCTGGGCGGTTCTGATAACGA | Construction of pET30a-LP-pcaIJFK |
| LP-HGBDC-REV | ACAAGGCCATGGCTGATCATATGAATATCCTCCTTAGTTGGCTTCAGGGATGAGGCGCCATCTTTGCCTGGCGGCAGTAG | Construction of pET30a-LP-pcaHGBDC |
| LP-control-REV | ACAAGGCCATGGCTGATCATATGAATATCCTCCTTAGTTGGCTTCAGGGATGAGGCGCCATCAAATAAATTTTTCATGATTT | Construction of pET30a-LP-control |
| ompT-LP-up | ATATAAAAAATACATATTCAATCATTAAAACGATTGAATGGAGAACTTTTCATATGAATATCCTCCTTAGTTGGCTTCAG | I-SceI, landing pad and CmR cassette integration at ompT |
| ompT-LP-down | AGTTATTCCCCGGGGCGATTTTCACCTCGGGGAAATTTTAGTTGGCGTTCGTGTAGGCTGGAGCTGCTTC | I-SceI, landing pad and CmR cassette integration at ompT |
| pflB-LP-up | TTTTACTGTACGATTTCAGTCAAATCTAATTACATAGATTGAGTGAAGGTGTGTAGGCTGGAGCTGCTTC | I-SceI, landing pad and CmR cassette integration at pflB |
| pflB-LP-down | CGAAGTACGCAGTAAATAAAAAATCCACTTAAGAAGGTAGGTGTTACATGCATATGAATATCCTCCTTAG | I-SceI, landing pad and CmR cassette integration at pflB |
| ompT-FWD | TAAACAAAATATAAACAGTGGAGC | PCR screen at ompT |
| ompT-REV | CTATAATTGATGGTTTTCTATGTG | PCR screen at ompT |
| pflB-FWD | TTTTACTGTACGATTTCAGTCAAAT | PCR screen at pflB |
| pflB-REV | CGAAGTACGCAGTAAATAAAAAATC | PCR screen at pflB |
| pTarget FWD | GTTTTAGAGCTAGAAATAGCAAGTTAAAATAAG | Inverse PCR of pTarget |
| pTarget REV | ACTAGTATTATACCTAGGACTGAG | Inverse PCR of pTarget |
| elfC N20F | TTCCTTATGCCACTTCTTATGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC | Construction of elfC sgRNA |
| elfC N20R | ATAAGAAGTGGCATAAGGAAACTAGTATTATACCTAGGACTGAGCTAGCTGTCAAGGATC | Construction of elfC sgRNA |
| pcaH N20F1 | TTTTACCTCCTCTCTAGAAGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC | Construction of pcaH sgRNA |
| pcaH N20R1 | CTTCTAGAGAGGAGGTAAAAACTAGTATTATACCTAGGACTGAGCTAGCTGTCAAGGATC | Construction of pcaH sgRNA |
| pcaH N20F2 | AGAGGAGGTAAAACATATGCGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC | Construction of pcaH sgRNA |
| pcaH N20R2 | GCATATGTTTTACCTCCTCTACTAGTATTATACCTAGGACTGAGCTAGCTGTCAAGGATC | Construction of pcaH sgRNA |
| flu N20F | TACGAACTTCCCTGTGCGGGGTTTTAGAGCTAGAAATAGCAAGTTAAAATAAGGCTAGTC | Construction of flu sgRNA |
| flu N20R | CCCGCACAGGGAAGTTCGTAACTAGTATTATACCTAGGACTGAGCTAGCTGTCAAGGATC | Construction of flu sgRNA |