Table 2 Oligonucleotides used in this study
Primers | Sequences | Notes |
|---|---|---|
aroG-F | CGCGCGTCGACATGAATAGGGGTGTGAGTTG | Amplification of aroG from genome, SalI and KpnI underlined |
aroG-R | CGCGCGGTACCTTAGTTACGCAGCATTTCTGCAACG | |
aroB-F | CGCGCGTCGACATGAGCGCAGTGCAGATTTTC | Amplification of aroB from genome, SalI and KpnI underlined |
aroB-R | CGCGCGGTACCTTAGTGGCTGATTGCCTCATAGCA | |
aroD-F | CGCGCGTCGACATGCCTGGAA AAATTCTCCT | Amplification of aroD from genome, SalI and KpnI underlined |
aroD-R | CGCGCGGTACCTTACTTTTTGAGATTTGCCAGGATA | |
aroE-F | CGCGCCTGCATATGGGTTCTCACATCACTCAC | Amplification of aroE from genome, PstI and KpnI underlined |
aroE-R | CGCGCGGTACCTTAGTGTTCTTCTGAGATGCCT | |
MU-aroG-1-F | GGCCTTACCGTTGGCAACATCAGCCAGCTTCTGCTTCAGCTCAAGTACC | Mutate HindIII in aroG |
MU-aroG-1-R | CCTGAGGTACTTGAGCTGAAGCAGAAGCTGGCTGATGTTG CCAACGGT | |
MU-aroG-2-F | TCGCGCCAACGTAAAGACTCTGCTCCAGATGGCAGTTGTTTTGACCT | Mutate PstI in aroG |
MU-aroG-2-R | CGTAGGTCAAAACAACTGCCATCTGGAGCAGAGTCTTTACGTTGGCGC | |
MU-aroG-3-F | GTGTCCGATGAGTCCCTGCGTGCTGCCGATATCTACTGCTCCCACGAGG | Mutate PstI in aroG |
MU-aroG-3-R | AGCCTCGTGGGAGCAGTAGATATCGGCAGCACGCAGGGACTCATCGGAC | |
MU-aroB-1-F | GCCTGACGCGGAAATCATCGCGGGTTCCGCCGAAATCATCAAAACTGG | Mutate BamHI in aroB |
MU-aroB-1-R | AACCAGTTTTGATGATTTCGGCGGAACCCGCGATGATTTCCGCGTCAGG | |
MU-aroB-2-F | CATCCGAGTTGGATGCAGCACTGGTCGCTGCTGGTTTGAAGGTCCTGC | Mutate SpeI in aroB |
MU-aroB-2-R | TGCAGGACCTTCAAACCAGCAGCGACCAGTGCTGCATCCAACTCGGATG | |
MU-aroD-F | TTAGCTCACCTTCGTGATTGCTCTGGAGCGCCTCAACCTCAAGGCCGTG | Mutate PstI in aroD |
MU-aroD-R | GCACGGCCTTGAGGTTGAGGCGCTCCAGAGCAATCACGAAGGTGAGCT | |
MU-aroE-2-F | CATGCCGTCTAAATTCGCAGCTCTTGAATTTGCCGACGAAGTAACCGAACGCGCCTGC | Mutate EcoRI in aroE |
MU-aroE-2-R | GCAGGCGCGTTCGGTTACTTCGTCGGCAAATTCAAGAGCTGCGAATTTAGACGGCATG | |
MU-aroE-2-F | ATGGCGCGCCGACAACACCGACGTTGACGGCATCAGGGGAGCTCTCGG | Mutate SalI in aroE |
MU-aroE-2-R | CACCGAGAGCTCCCCTGATGCCGTCAACGTCGGTGTTGTCGGCGCGCC | |
RiboJ-F | CGCGAAGCTTAGCTGTCACCGGATGTGCTTTCCGGTCTGATGAGTC | Amplification of RiboJ from pUC19, HindIII and PstI underlined |
RiboJ-R | CGCGCTGCAGTTAAACAAAATTATTTGTAGAGGCTGTTTCG | |
EGFP-F | CGCGGGTACCGTGAGCAAGGGCGCCGAGC | Amplification of egfp from pACGFP, KpnI and EcoRI underlined |
EGFP-R | CGCGGAATTCTCACTTGTACAGCTCATCCATGCCGTGGGT | |
MU-RBSAG-F | CGCGCGTCGACAAAGGNNNNNNNNATGAATAGGGGTGTGAGTTG | Amplification of aroG with mutated RBS, SalI and KpnI underlined |
MU-RBSAG-R | CGCGCGGTACCGTTACGCAGCATTTCTGCAACG | |
MU-RBSAB-F | CGCGCGTCGACAAAGGNNNNNNNNATGAGCGCAGTGCAGATTTTC | Amplification of aroB with mutated RBS, SalI and KpnI underlined |
MU-RBSAB-R | CGCGCGGTACCGTGGCTGATTGCCTCATAAGCA | |
MU-RBSAD-F | CGCGCGTCGACAAAGGNNNNNNNNATGCCTGGAAAAATTCTCCT | Amplification of aroD with mutated RBS, SalI and KpnI underlined |
MU-RBSAD-R | CGCGCGGTACCCTTTTTGAGATTTGCCAGGATA | |
MU-RBSAE-F | CGCGCGTCGACAAAGGNNNNNNNNATGGGTTCTCACATCACTCAC | Amplification of aroE with mutated RBS, SalI and KpnI underlined |
MU-RBSAE-R | CGCGCGGTACCGTGTTCTTCTGAGATGCCT | |
aroG-H-F | CGCGCGTCGACAAAGGGTGAATCTATGAATAGGGGTGTGAGTTG | aroG with high strength RBS, SalI and BamHI underlined |
aroG-H-R | CGCGCGGATCCTTAGTTACGCAGCATTTCTGCAACG | |
aroG-M-F | CGCGCGTCGACAAAGGTTCTAAAGATGAATAGGGGTGTGAGTTG | aroG with medium strength RBS, SalI and BamHI underlined |
aroG-M-R | CGCGCGGATCCTTAGTTACGCAGCATTTCTGCAACG | |
aroG-L-F | CGCGCGTCGACAAAGGGCCGAATTATGAATAGGGGTGTGAGTTG | aroG with lows trength RBS, SalI and BamHI underlined |
aroG-L-R | CGCGCGGATCCTTAGTTACGCAGCATTTCTGCAACG | |
aroB-H-F | CGCGCGTCGACAAAGGGGAGAGCCATGAGCGCAGTGCAGATTTTC | aroB with high strength RBS, SalI and BamHI underlined |
aroB-H-R | CGCGCGGATCCTTAGTGGCTGATTGCCTCATAAGCA | |
aroB-M-F | CGCGCGTCGACAAAGGCATGTTCTATGAGCGCAGTGCAGATTTTC | aroB with medium strength RBS, SalI and BamHI underlined |
aroB-M-R | CGCGCGGATCCTTAGTGGCTGATTGCCTCATAAGCA | |
aroB-L-F | CGCGCGTCGACAAAGGAACGACTAATGAGCGCAGTGCAGATTTTC | aroB with low strength RBS, SalI and BamHIunderlined |
aroB-L-R | CGCGCGGATCCTTAGTGGCTGATTGCCTCATAAGCA | |
aroD-H-F | CGCGCGTCGACAAAGGAGGTTGTCATGCCTGGAAAAATTCTCCT | aroD with high strength RBS, SalI and BamHI underlined |
aroD-H-R | CGCGCGGATCCTTACTTTTTGAGATTTGCCAGGATA | |
aroD-M-F | CGCGCGTCGACAAAGGCATGGCCGATGCCTGGAAAAATTCTCCT | aroD with medium strength RBS, SalI and BamHI underlined |
aroD-M-R | CGCGCGGATCCTTACTTTTTGAGATTTGCCAGGATA | |
aroD-L-F | CGCGCGTCGACAAAGGTGGTTCATATGCCTGGAAAAATTCTCCT | aroD with low strength RBS, SalI and BamHI underlined |
aroD-L-R | CGCGCGGATCCTTACTTTTTGAGATTTGCCAGGATA | |
aroE-H-F | CGCGCGTCGACAAAGGAGGATTAGATGGGTTCTCACATCACTCAC | aroE with high strength RBS, SalI and BamHI underlined |
aroE-H-R | CGCGCGGATCCTTAGTGTTCTTCTGAGATGCCT | |
aroE-M-F | CGCGCGTCGACAAAGGAGAACGTGATGGGTTCTCACATCACTCAC | aroE with medium strength RBS, SalI and BamHI underlined |
aroE-M-R | CGCGCGGATCCTTAGTGTTCTTCTGAGATGCCT | |
aroE-L-F | CGCGCGTCGACAAAGGGTTCATAGATGGGTTCTCACATCACTCAC | aroE with low strength RBS, SalI and BamHI underlined |
aroE-L-R | CGCGCGGATCCTTAGTGTTCTTCTGAGATGCCT | |
PB-F | CGCGGGATCCTTGCGCCGACATCATAACGGTT | BamHI and XmaI underlined |
PB-R | CGCGCCCGGGTTAGTGGCTGATTGCCTCATAAGCA | |
PD-F | CGCGCCCGGGTTGCGCCGACATCATAACGGTT | XmaI and KpnI underlined |
PD-R | CGCGGGTACCTTACTTTTTGAGATTTGCCAGGATA | |
PE-F | CGCGGGTACCTTGCGCCGACATCATAACGGTT | KpnI and EcoRI underlined |
PE-R | CGCGGAATTCTTAGTGTTCTTCTGAGATGCCT | |
Terminator 1-F | CGCGCCCGGGGGCTGTTTTGGCGGATGAGAGAAGATTTTC | XmaI underlined |
Terminator 1-R | CGCGCCCGGGAGAGTTTGTAGAAACGCAAAAAGGCC | |
Terminator 2-F | CGCGGGTACCGGCTGTTTTGGCGGATGAGAGAAGATTTTC | KpnI underlined |
Terminator 2-R | CGCGGGTACCAGAGTTTGTAGAAACGCAAAAAGGCC | |
Terminator 3-F | CGCGGGATCCGGCTGTTTTGGCGGATGAGAGAAGATTTTC | BamHI underlined |
Terminator 3-R | CGCGGGATCCAGAGTTTGTAGAAACGCAAAAAGGCC | |
aroK-F | CGCGGAATTCTGGCTGATTGCCTCATAAGCACTCT | EcoRI and HindIII underlined |
aroK-R | CGCGAAGCTTTTCGATGGACTACAGCAGGTGAATC | |
KTaroK-F | CGCGCCCGGGCCTCTAAACCTTCGAATTTCATTCGTTCCTC | XmaI underlined |
KTaroK-R | CGCGCCCGGGCGATTAATTAAACCGGGCACCTGATTAAC | |
V-KTaroK-F | TCCATGCTGGGCTGGTGCAAAATCGCTACC | Primer used to verify ΔaroK |
V-KTaroK-R | AACCATTGATATGGAAAACGGCAAGGCAGC |