Table 2.
Primers used for cloning and construction of unmarked gene deletions.
| Name | Sequence | Restriction site |
|---|---|---|
| expfor_Chol1_3αHSD | TTTTTTGGATCCATGTCCGTTATCGCAATTA | BamHI |
| exprev_Chol1_3αHSD | TTTTTTGAATTCTCAGAAGACCTTGGTCCGCA | EcoRI |
| expfor_Chol1_c211_11427 | TTTTTTTCTCGAGATGCCCGACTATTCGCACCT | XhoI |
| exprev_Chol1_c211_11427 | TTTTTTTTCTAGATCAAACCCTCAACGCCACCT | XbaI |
| expfor_RHA1_casH | TTTTTTTAAGCTTATGACGATAGATCTCGACCCGCC | HindIII |
| exprev_RHA1_casH | TTTTTTTTCTAGATCACGGCACGCGAAGCAC | XbaI |
| expfor_Chol11_5β-Δ4-kstd1 | TTTTTTTCTCGAGATGACTGCTGCCCCTGCCCT | XhoI |
| exprev_Chol11_5β-Δ4-kstd1 | TTTTTTTTCTAGATCAGCGGCCCATGGCGG | XbaI |
| expfor_Chol11_nov2c085 | TTTTTTTCTCGAGATGGCTTACCGCCACCTCCT | XhoI |
| exprev_Chol11_nov2c085 | TTTTTTTTCTAGATCATCCGATCGCGGCGACGG | XbaI |
| expfor_Chol11_nov2c314 | TTTTTTTCTCGAGATGGAATTGCGCAACCGGAT | XhoI |
| exprev_Chol11_nov2c314 | TTTTTTTTCTAGATCACCCGGCTATTGCGTCCA | XbaI |
| purfor_Chol11_5β-Δ4-kstd1 | TTTTTTTCATATGACTGCTGCCCCTGCCCT | NdeI |
| purrev_Chol11_5β-Δ4-kstd1 | TTTTTTTCTCGAGTGAGCGGCCCATGGCGG | XhoI |
| upfor_Chol11_5β-Δ4-kstd1 | TTTTTTTTCTAGAGTAGAGGTGGAGCGGAATC | XbaI |
| uprev_Chol11_5β-Δ4-kstd1 | TCTGATCTTCAAGGCCAAAG | |
| dnfor_Chol11_5β-Δ4-kstd1 | CTTTGGCCTTGAAGATCAGAGAGAGTCTCCTAATTCGCCA | |
| dnrev_Chol11_5β-Δ4-kstd1 | TTTTTTTCTCGAGCTACGACAACCCGGTCAACA | XhoI |
| upfor_Chol1_c211_11427 | TTTTTTTTCTAGATGGGACGGGACCAAATTACG | XbaI |
| uprev_Chol1_c211_11427 | GGTAGGTGGATGAAAAAGAAAGCC | |
| dnfor_Chol1_c211_11427 | TTCTTTTTCATCCACCTACCGTTTCGTTCCTTGCGATGGGG | |
| dnrev_Chol1_c211_11427 | TTTTTTTCTCGAGCGGGTCGAAAAGCTGATCCT | XhoI |
| upfor_Chol1_stdA1 | TTTTTTTTCTAGACTCACGCAATCACGCCACTC | XbaI |
| uprev_Chol1_stdA1 | CTGCCGTTCCCCTGTCACCA | |
| dnfor_Chol1_stdA1 | TGGTGACAGGGGAACGGCAGCAGCGCCTCACGCAGCGGCG | |
| dnrev_Chol1_stdA1 | TTTTTTTCTCGAGGACGTGGTCCCAGCCAGGCA | XhoI |
| upfor_Chol1_kstd1 | TTTTTTTCTAGAACCAACAACTCCAGCCTCG | XbaI |
| uprev_Chol1_kstd1 | GTCCGACTCACTTGCCAGGTT | |
| dnfor_Chol1_kstd1 | AACCTGGCAAGTGAGTCGGACGGAGCGACCGATGAATGCACG | |
| dnrev_Chol1_kstd1 | TTTTTTAAGCTTATGGAGGTAACCGAGGCGA | HindIII |
| M13 for (−43) | AGGGTTTTCCCAGTCACGACGTT | |
| M13 rev (−49) | GAGCGGATAACAATTTCACACAGG | |
| pDM4_MCS_for | ACTTAACGGCTGACATGGGA | |
| pDM4_MCS_rev | GCGAAGTGATCTTCCGTCAC | |
| T7_for | TAATACGACTCACTATAGGG | |
| T7_rev | TATGCTAGTTATTGCTCAG | |
| pDM4_backbone_for | AAG ATG TGG CGT GTT ACG GT | |
| pDM4_backbone_rev | AGG CTC TGG GAG GCA GAA TA | |
| sacB_for | AGGAGACATGAACGATGAACA | |
| sacB_rev | TTTTTTTCCCGGGTCGGCATTTTCTTTTGCGTT |
Underlines indicate restriction sites.