Table 1.
Primers for upstream, downstream, and internal regions used to construct deletion strainsa
| Target | Forward primer (3′→5′) | Reverse primer (5′→3′) |
|---|---|---|
| or0897 | ||
| Upstream | AGGAGAATTTGCAAGAATGA AGG | CAATTTACCTCCTCTCCGCT G |
| Downstream | GAAGGCATTTAATGGTTTTC AGC | CCAACACCATTCTCTTACAT TACCC |
| Internal | GATGTTAAACTTTAAGAGAA TTTTTACGTTAATTTG | CTCACCAAACTGTTACATTA GCATAACC |
| or1451 | ||
| Upstream | CGATGGAGAATTTAAATTTG CAGCC | TTTAAAACACCTCTTCTTAA ATAAATTTCATTCAACC |
| Downstream | ACTTTGGGGGTCTTAACATG ATTAAAG | CCATGAAATTGCGATCGC |
| Internal | GGAGCTTTTAAAGAGAGACG ATGG | ACACCTGTTGCCACATTGC |
| or1452 | ||
| Upstream | GTCTTCTGGGAAATGAGGTT CC | AAAGTCTATTTCATCCACAA TACACTTGTC |
| Downstream | TATGCATTTGGAGGGATTGA TATG | TAATGTCGTAGGCATACAAG ACCC |
| Internal | CAGGCAGACTTGGCCTTG | CATATGCTCATCTCTATAAA GCATTTCC |
| or1456 | ||
| Upstream | CAAGCCTATCATACCATTTA GCTGTC | ACAAGATATTAAAAAATTCA GAAAATATCAGAAATATTT |
| Downstream | TGAATTAATATAATTAAAAA CTTTCTGTAATGAAGTACG | AAGAAACAACTGAGTTAAAT ATACCGGC |
| Internal | AAAACTTTCTGAATTGACAG GTGTG | GAATAATCTTTACCTTCAAT TCCCCAG |
| or1457 | ||
| Upstream | TAATAAAGCACATCCATTTG CAATAAG | AAATAATATTTTAAGCGTAA TGAAGTACGGAAC |
| Downstream | TCTAATTTACAAGATTTCAG GGGGTTATAAAGTGGC | TGTCTAATAAGTATTTTTAA ATTTCCCCCC |
| Internal | CATGGCTGTTACAGATGCTA GAGTAG | TTAAAATGCACTTTCACCAC TAAGTG |
| or1458 | ||
| Upstream | GATGCTAGAGTAGAAGTTTT ATCAGAAGGG | TTTATAACCCCCTGAAATCT TGTAAATTAG |
| Downstream | TACATCTTTGGTTTTTTATA AATGCTGAAAG | CCATTCGCCTTTATTTACAT TTGC |
| Internal | GTGGCTAAAAATAATAGAAT AAGGTTGAGC | CTAACCCTTAACACTGCCAA TGG |
| or1459 | ||
| Upstream | AGCACAGAAGATATTGTATT TGACACG | TTCTTACTTCCTCCCTCAGT AAATTTAATTTATTG |
| Downstream | AAAAAACAAATAATCTTTAA GTAAAAAGGCAG | AAAAAGGCTTCAGAATGGCT TG |
| Internal | GAACATTATATAGTTAAGAG GGATGGTACAGG | CACAACTTTCTGTTGTCCTT CACC |
| Endoxylanase region | ||
| Upstream | CATGATTCCGGCAGGC | CGCAACATATCTACCCATAT ATACCATC |
| Downstream | GCCTGTAAATTCGAATTCAC CTG | GCCAATGGTGAATTGAAAGA AAATG |
| Internal | CAATGCTTTTTTGTTCCATG AATC | CTTGTATTGGATTTTGTGGT GTCACTAC |
| β-Xylosidase region | ||
| Upstream | AAAGTCTGTGGCGATGGAGA | GCATTTGTTTGAGCTTCCA |
| Downstream | GGAGATGCTTTAAGCATCTC CT | TGGGTGAAGAATCTCGCTTG |
| Internal | GGGACTTTTCGACATGCCAT TAC | CCAACGCACATTAAGACATC G |
| Xylose catabolism region | ||
| Upstream | CACCGTTTTCTACGTAGCTT CTGGCT | GGCATGTAAGTCCTTCCTCG C |
| Downstream | AGCTTCCTCCTTCTATTCGT TTGTCTG | GGTCCTGCTGGAATGTCTGT TG |
| Internal | ATCCAACTTCCCAATTTCTT CAAAGG | CCAGAGGACACAGGAAGAAT GC |
| pta/ack/kan cassette | CGTGCCCATTGTGAAGTGG | GCGCCTACGAGGAATTTGTA TCG |
a
Overlapping sequences used in vector construction, matching an adjacent region, are not shown.