Table 2. Strains, plasmids and primers used in this study.
| Strain | Genotype | Reference |
|---|---|---|
| Escherichia coli DH5α | F− λ−endA1 glnV44 thi-1 recA1 relA1 gyrA96 deoR nupG Φ80 lacZ ΔM15 Δ(lacZYA-argF) U169, hsdR17(rK− mK+) | Invitrogen |
| E. coli ER2566 | F− λ−fhuA2 [lon] ompT lacZ::T7 gene 1 gal sulA11 Δ(mcrC-mrr)114::IS10 R(mcr-73::miniTn10-TetS)2 R(zgb-210::Tn10)(TetS) endA1 [dcm] | NEB |
| E. coli W3110 | F− λ−rph-1 INV(rrnD, rrnE) | (39) |
| Plasmid | Description | Source |
| pALO3 | cptNEr in pTYB1 | This study |
| pALO5 | cptIEr with PT7 upstream in pACYC184 | This study |
| pFLS104 | cptIRt repeats and terminator in pTA100, SpR | (16) |
| pFLS105 | cptNRt in pBAD30, ApR | (16) |
| pFLS106 | cptICc repeats and terminator in pTA100, SpR | (16) |
| pFLS107 | cptNCc in pBAD30, ApR | (16) |
| pFLS108 | cptIEr repeats and terminator in pTA100, SpR | (16) |
| pFLS109 | cptNEr in pBAD30, ApR | (16) |
| pFLS110 | cptIEr single repeat in pTA100, SpR | This study |
| pFLS111 | cptIEr ΔU30 single repeat in pTA100, SpR | This study |
| pFLS112 | cptIEr A29U single repeat in pTA100, SpR | This study |
| pFLS122 | cptINEr locus in pRBJ200, ApR | This study |
| pRBJ200 | E. coli par-deficient single-copy vector, ApR | (40) |
| pTA100 | SpR derivative of pQE-80L | (13) |
| Primer | Sequence 5′-3prime;* (Restriction site) | |
| CptI-WT | tttaagcttTCAACCCGACCATTTATATACCACATATCGGTCAGTGGTAAACTTtcaattgaatctattataattgtta (HindIII) | |
| CptI-dU30 | tttaagcttTCAACCCGACCATTTTATACCACATATCGGTCAGTGGTAAACTTtcaattgaatctattataattgtta (HindIII) | |
| CptI-A29U | ttaagcttTCAACCCGACCATTTAAATACCACATATCGGTCAGTGGTAAACTTtcaattgaatctattataattgtta (HindIII) | |
| PF185 | AAACAAATAGGGGTTCCG | |
| TRB271 | TTGGATCCATAATCAGTATCACTGAGAAA (BamHI | |
| TRB272 | TTTCTAGAACTACGGCAAAAGACTTTTTC (XbaI) | |
| TRB273 | CCGGCATATGATAAGGAATGGTTTTTATATTATC (NdeI) | |
| TRB274 | GTGGTTGCTCTTCCGCACAAATGTGTTGCTGGTAAATC (SapI) |
*For CptI primers, the region encoding the repeat is shown in uppercase.