Table 1 Construction of gene knockout and complementation plasmids.
Plasmid name | Primer position | Primer Sequence 5'-3' | Number of 5' codons | Number of 3' codons |
|---|---|---|---|---|
pBBΔorc1 | 5' forward | CGCAAGCTTGACTCCACCCTTCCGAGAGT | 3 | 2 |
| Â | 5' reverse | CGCACTAGTGGTGATCATGGGTTTGCGTC | Â | Â |
| Â | 3' forward | CGCACTAGTCGAAACTAGCTCTCCAAGCTC | Â | Â |
| Â | 3' reverse | CGCGGATCCTCTACTGTACAGCAGATGAG | Â | Â |
pBBΔorc2 | 5' forward | CGCAAGCTTCAACAAAATTATGCGTAGAG | 2 | 3 |
| Â | 5' reverse | CGCACTAGTCGTCATTGAATATCACACGG | Â | Â |
| Â | 3' forward | CGCACTAGTGCCACGTTCTGAGTATTCTG | Â | Â |
| Â | 3' reverse | CGCGGATCCGATCTTGTACTTGTCCTCGC | Â | Â |
pBBΔorc3 | 5' forward | CGCGGATCCTGAAACGGGTCTGTGAGTGG | 4 | 6 |
| Â | 5' reverse | CGCACTAGTCCCTTTCACCATCTCGATAA | Â | Â |
| Â | 3' forward | CGCACTAGTCAGACGTTGATGGGATGGTGA | Â | Â |
| Â | 3' reverse | CGCGAATTCGGTGAGGACCTCGAGTTCGAT | Â | Â |
pBBΔorc4 | 5' forward | CGCAAGCTTCAGGAGACAGCTACCTACGA | 3 | 2 |
| Â | 5' reverse | CGCACTAGTGGGCGTCATTGAATATCACA | Â | Â |
| Â | 3' forward | CGCACTAGTCGCGAGTAATGACCCCTACT | Â | Â |
| Â | 3' reverse | CGCGGATCCTCGGCTATCAAGGGTTCAGC | Â | Â |
pBBΔorc5 | 5' forward | CGCAAGCTTAGTTGGTGACGCTCATCGGC | 4 | 2 |
| Â | 5' reverse | CGCGAATTCGCGACCAGCCATACGTATGC | Â | Â |
| Â | 3' forward | CGCGAATTCGAGGAGTAGTTGCCAGGCGA | Â | Â |
| Â | 3' reverse | CGCGGATCCTGATCGAGGCGAACACGCTG | Â | Â |
pBBΔorc6 | 5' forward | CCGGAATTCTTCGAGGCGACGCTGCGGGA | 7 | 6 |
| Â | 5' reverse | CTCTTCGGGGTCCTCATCCAT | Â | Â |
| Â | 3' forward | GCGGTCCTGGAGCGCCTGTAG | Â | Â |
| Â | 3' reverse | CCGGAATTCCAGCGCCTCAACCCGATCGAC | Â | Â |
pBBΔorc7 | 5' forward | CGCGGATCCGCGCCCGAACGCAACTAGAA | 3 | 3 |
| Â | 5' reverse | GCGCTCGAGGTCTGTCATGTATTCACGCAC | Â | Â |
| Â | 3' forward | GCGCTCGAGGAGAACAATTAGTGGATGCT | Â | Â |
| Â | 3' reverse | GCGAAGCTTGGTGTGATGTTCATGACCAT | Â | Â |
pBBΔorc8 | 5' forward | CCGGAATTCCACGTCGTGTTGGCGGTGGT | 3 | 3 |
| Â | 5' reverse | GCGCTCGAGCGTCTTCATCGCTTGCCGAGA | Â | Â |
| Â | 3' forward | GCGCTCGAGGCGACCGTGTAGACCCCGGA | Â | Â |
| Â | 3' reverse | CGCAAGCTTGATGAAGCTCCGCCGCAGCG | Â | Â |
pBBΔorc9 | 5' forward | CGCAAGCTTTGGGTCGTGTACACGGCCTC | 4 | 3 |
| Â | 5' reverse | CGCACTAGTGCGGCAGGTCATATAAGAGT | Â | Â |
| Â | 3' forward | CGCACTAGTGTAACCCAATAAGCTGCGAA | Â | Â |
| Â | 3' reverse | CGCGGATCCAACACTCATCGACGAGTGAA | Â | Â |
pBBΔorc10 | 5' forward | CCGGATCCCCTGTGGTCGTTCTGGAAGAC | 5 | 2 |
| Â | 5' reverse | CCACTAGTACCCAACAGTGACATCCTCC | Â | Â |
| Â | 3' forward | CGCACTAGTGTGCTGTAGCGATTGTGCGA | Â | Â |
| Â | 3' reverse | CGCCTCGAGCACGGAGACGTCGAGAGCGAG | Â | Â |
pBBΔpolD1 | 5' forward | CCGGAATTCGTCGGTCGGATCGGGGACAT | 2 | 2 |
| Â | 5' reverse | GCGCTCGAGCGTCATGTCCCCGTCGATCT | Â | Â |
| Â | 3' forward | GCGCTCGAGTTCTCGTAGCCACGGCGGCG | Â | Â |
| Â | 3' reverse | GCGAAGCTTCGTTGTGCACCATCGTCACC | Â | Â |
pBBΔpolD2 | 5' forward | GCGGAATTCGGCGTGGCGGTAACGGCGTT | 1 | 2 |
| Â | 5' reverse | GCGACTAGTCATTACAGCCAGCGGTCGAG | Â | Â |
| Â | 3' forward | GCGACTAGTTTCATGTGACCACGGCGCTC | Â | Â |
| Â | 3' reverse | GCGAAGCTTGACGTGATCGACGAACACACC | Â | Â |
pBBΔpolB1 | 5' forward | CCGGAATTCGCCAACACTGCCGCGTTGAA | 3 | 3 |
| Â | 5' reverse | CGCACTAGTGTTTCCCATTGGGTTCGGGT | Â | Â |
| Â | 3' forward | CGCACTAGTCAGTTCACGTAGCCCGCTGG | Â | Â |
| Â | 3' reverse | CGCAAGCTTCTCGTCAACAACGCCGGGCT | Â | Â |
pBBΔpolB2 | 5' forward | CGCGAATTCCCGAACGACGCGGCATCAAG | 2 | 2 |
| Â | 5' reverse | CGCACTAGTCGGCATTCCCTACAGAACCA | Â | Â |
| Â | 3' forward | CGCACTAGTCTGTCGTAGTGGACCCCACC | Â | Â |
| Â | 3' reverse | CGCAAGCTTCGGCTTTTCCTGGCCAAGTC | Â | Â |
pBBΔmcm | 5' forward | CGCGAATTCGTCGAGAACCCCAGGATGAG | 3 | 3 |
| Â | 5' reverse | CGCACTAGTCGGATCCATCTGGTAGAGAT | Â | Â |
| Â | 3' forward | CGCACTAGTCGCTCGATCTAGCCGACGGC | Â | Â |
| Â | 3' reverse | CGCAAGCTTACAGCACGCCCACGTGCTCGT | Â | Â |
pBBΔpri1 | 5' forward | ATGAGTCCCCCACTCGGTCTT | 2 | 6 |
| Â | 5' reverse | GTGCATGCCGGCAATCGTGG | Â | Â |
| Â | 3' forward | GCCGAGAAGGCCACAGAATGA | Â | Â |
| Â | 3' reverse | GGCGGCTCTTCGACCTGACT | Â | Â |
pBBΔpri2 | 5' forward | TCTGTCAGGACCGGGCCACT | 2 | 2 |
| Â | 5' reverse | GTTCATGCTGGCCCGTGTTTG | Â | Â |
| Â | 3' forward | CCCGATTAGCCCTGCTTGCC | Â | Â |
| Â | 3' reverse | ATGGCTAACTCCAACGCCAA | Â | Â |
pBBΔpcn | 5' forward | TCTCGTCTGCGGCGGGGGTA | 4 | 4 |
| Â | 5' reverse | CGCCTTGAACATTATTGCAGA | Â | Â |
| Â | 3' forward | ATCCAGTCCAACTGACGCCA | Â | Â |
| Â | 3' reverse | ATGCTGGCCCGTGTTTGCGA | Â | Â |
pBBΔrad2 | 5' forward | CCGGAATTCGTGATGTCGAACACGGGGAA | 3 | 2 |
| Â | 5' reverse | CGCCTCGAGGTTCCCCATCACGCCCAGTT | Â | Â |
| Â | 3' forward | CGCCTCGAGTGGACGTGACCGGCGCTGAT | Â | Â |
| Â | 3' reverse | CGCAAGCTTGCGTAAAAGCCATCGGAACC | Â | Â |
pBBpolD1 all | 5' forward | CGCGAATTCCATACCGGTTCGGTCGTACC | Â | Â |
| Â | 3' reverse | CGCACTAGTCAGGAAGTCCTCTCGCATAC | Â | Â |
pBBpolB1 all | 5' forward | CGCGAATTCGTCACCTCGGTCCGTGGTAG | Â | Â |
| Â | 3' reverse | CGCACTAGTGCGTTCGCGGCGACCCAGAGT | Â | Â |
pBBmcm all | 5' forward | CGCGAATTCGAACAGCATGAACATGCCGA | Â | Â |
| Â | 3' reverse | CGCACTAGTCGAATACGCCACTGCTAACAA | Â | Â |
pBBpri2 all | 5' forward | CGCGAATTCGGCGTACTTCCACGTCCAGGG | Â | Â |
| Â | 3' reverse | CGCACTAGTGCCACGTCCGGGTACGCAGTG | Â | Â |
pBBrad2 all | 5' forward | CGCGAATTCCCAGCACGAGTCGAGTGGTAA | Â | Â |
| Â | 3' reverse | CGCACTAGTATCCATGCCTGTGCGTGAGC | Â | Â |