Table 2.
Oligonucleotides used in this study
| Oligonucleotide | Sequence (5′–3′) | Restriction site |
|---|---|---|
| del_cg3349-54-up-s | ACCAAGCTTCATTTTCTAGTTGAGATTCATTATCATAACGCTTGACAGTACAACTATGTC | HindIII |
| del_cg3349-54-up-as | GCTTCCGGGGTGAATGACGGAGCCGCTGCGGAGGTTCCAGAGCCACCGCGCTG | – |
| del_cg3349-54-down-s | CGCAGCGGCTCCGTCATTCACCCCGGAAGCTGGCGCTACGCCCTCCTCGAAC | – |
| del_cg3349-54-down-as | ACCTCTAGATTTCTGTCTTGAGGGTTCGTGGGGGCTG | XbaI |
| check-cg3349-54-s | AAAGGCTCCATGAATTCCTCACGGAGGATCTC | – |
| check-cg3349-54-as | ACATCACAAGTAGAAACCCGCATTTTCTGTAGTTTTTAC | – |
| (A)-aroF*-s | TTCGCTCTTCAAAGCTGCTTAAGGAGGCTATCTATGACCGATGAACAGGTGCTGATGACCCCAG | SapI |
| (A)-aroF*-as | TACGCTCTTCTGATTTATGCCACGCGTGCGGTCAGCTG | SapI |
| (A)-aroH-s | TTCGCTCTTCAAAGTATACCATGGTAAGGAGGTTCAGCATGAACAGAACTGACGAACTCCGTACTGCGCGTATTG | SapI |
| (A)-aroH-as | TACGCTCTTCTGATTTAGAAGCGGGTATCTACCGCAGAGGCGAGTTTTTC | SapI |
| (B)-qsuB-s | TTCGCTCTTCAATCTCTATCAAGGAGGATCGGCATGCGTACATCCATTGCCACTGTTTGTTTGTC | SapI |
| (B)-qsuB-as | TACGCTCTTCTTCGTTAGTTTGGGATTCCCCGCTCGAGGTC | SapI |
| (B)-irp9-s | TTCGCTCTTCAATCTCTATCAAGGAGGATCGGCATGAAGATCTCCGAGTTCCTCCACCTGGCAC | SapI |
| (B)-irp9-as | TACGCTCTTCTTCGTTACACCATCAGGTATGGTGCAATGGAGGCCAGCTTTTCGCGAGTTTCG | SapI |
| (B)-hyg5-s | TTCGCTCTTCAATCTCTATCAAGGAGGATCGGCATGAACCCATCCTCCTTGGTGCTGAAC | SapI |
| (B)-hyg5-as | TACGCTCTTCTTCGTTACATGACCACGCCTTCGATTTCCACGAG | SapI |
| (B)-ubiC-s | TTCGCTCTTCAATCTCTATCAAGGAGGATCGGCATGTCACACCCCGCGTTAACGCAACTG | SapI |
| (B)-ubiC-as | TACGCTCTTCTTCGTTAGTACAACGGTGACGCCGGTAAAAACAGTTCTGTTAG | SapI |
| tkt’-s | CATGGATCCAAGGAGGTTCAGCATGACCACCTTGACGCTGTCACCTGAACTTCAG | BamHI |
| tkt’-as | AGAGCTCTTCAGCCCATAGCGTGCTCACGGATACCGAAGTG | SapI |
| ‘tkt-s | TTTGCTCTTCTGGCTCCATCCTCAACGGCATTTCCCTCC | SapI |
| ‘tkt-as | TCTGAATTCTTAACCGTTAATGGAGTCCTTGGCCGCTGCCAC | EcoRI |
| pMKEx2_own-s | CCCTCAAGACCCGTTTAGAGGC | – |
| pMKEx2_own-as | TTAATACGACTCACTATAGGGGAATTGTGAGC | – |
| pEKEx3-s | GCAAATATTCTGAAATGAGCTGTTGACAATTAATCATC | – |
| pEKEx3-as | CGTTCTGATTTAATCTGTATCAGGCTGAAAATCTTCTC | – |
Restriction sites are underlined; SapI cuts outside of its recognition site, the obtained 5′-overhangs after SapI cleavage used for Electra Cloning are shown in bold. In case of native tkt two separate PCR fragments were assembled using Electra Cloning to eliminate an internal SapI restriction site