TABLE 2.
Plasmids
| Plasmid | Featuresa | Construction or referenceb |
|---|---|---|
| pBAD30 | araC Para MCS in p15A-ori; Ampr | 38 |
| pCP20 | cI857 PR flp in pSC101-repts Ampr | 39 |
| pKD3 | FRT-cmR-FRT in oriRγ Ampr | 33 |
| pKD4 | FRT-kanR-FRT in oriRγ Ampr | 33 |
| pKD46 | araC Para γ-β-exo in pSC101-repts Ampr | 33 |
| pSUB11 | 3xFLAG in pKD4 | 34 |
| pKESK22 | lacIq Ptac MCS in p15A-ori; Kanr | 40 |
| pKETS24 | lacI PlacUV5 in pSC-ori Cmr | 41 |
| pKESL165 | lacIq Ptac mVenus in pKESK22 | 9 |
| pKESL166 | lacIq Ptac pdeL-mVenus in pKESK22 | 9 |
| pKESL209 | lacIq Ptac pdeLHTH5M-mVenus in pKESK22 | 9 |
| pKESL210 | lacIq Ptac pdeLEVL-AAA-mVenus in pKESK22 | pdeLEVL-AAA (flanking oligonucleotides T925/T972, mutagenesis oligonucleotides OA167/OA168) in pKESL165 |
| pKEHB12 | 3×FLAG in pBAD30 | 3×FLAG (annealed oligonucleotidess T687/T906) in pBAD30 |
| pKEHB23 | araC Para mVenus in pBAD30 | mVenus (pKESL165 EcoRI/XbaI) in pBAD30 |
| pKECY1 | araC Para pdeL-mVenus in pBAD30 | pdeL (T925/T952) in pKEHB23 |
| pKECY11 | araC Para pdeLHTH5M-mVenus in pBAD30 | pdeLHTH5M (T925/T952 from pKESL209) in pKEHB23 |
| pKECY15 | mCherry in pKD4 | mCherry (PCR OA480/OA481) in pKD4 |
| pKECY26 | mNeonGreen in pSC-ori Cmr | 9 |
| pKECY38 | mNeonGreen in pKD3 | mNeonGreen (SalI, BamHI from pKECY26) |
| pKECY43 | ftsZ-mNeonGreen in in pSC-ori Cmr | ftsZ (PCR OA807/OA808) in pKECY26 (XbaI, NdeI) |
| pKECY44 | araC Para pdeL in pBAD30 | 9 |
| pKECY52 | araC Para pdeLHTH5M in pBAD30 | 9 |
| pKECY53 | araC Para pdeLEVL-AAA in pBAD30 | 9 |
| pKECY81 | araC Para pdeL-3×FLAG in pBAD30 | pdeL (T925/OA116) in pKEHB12 |
| pKECY90 | lacI PlacUV5 ftsZ-mNeonGreen in pSC-ori Cmr | ftsZ-mNeonGreen (pKECY43 NcoI, XbaI) in pKETS24 |
| pKECY91 | araC Para pdeLHTH5M-3×FLAG in pBAD30 | pdeLHTH5M (PCR T925/OA116, pKECY52) in pKEHB12 |
| pKECY92 | araC Para pdeLEVL-AAA-3×FLAG in pBAD30 | pdeLEVL-AAA (PCR T925/OA116, pKECY53) in pKEHB12 |
| pKECY95 | araC Para rcsB-3×FLAG (native-RBS) in pBAD30 | rcsB (PCR T358/S866) in pKEHB12 |
| pKECY96 | araC Para rcsB-3×FLAG (T7gene10-RBS) in pBAD30 | rcsB (PCR OB93/S866) in pKEHB12 |
| pKECY97 | araC Para rcsB-3×FLAG (pdeL-RBS) in pBAD30 | rcsB (PCR OB94/S866) in pKEHB12 |
| pKECY98 | araC Para rcsB (T7gene10-RBS) in pBAD30 | rcsB (PCR OB93/T106) in pBAD30 |
| pKECY99 | araC Para lacI (T7gene10-RBS) in pBAD30 | lacI (PCR OB155/OB156) in pKECY98 |
| pKECY101 | araC Para rutR (T7gene10-RBS) in pBAD30 | rutR (PCR OB159/OB160) in pKECY98 |
| pKECY102 | araC Para leuO (T7gene10-RBS) in pBAD30 | leuO (PCR OA005/T558) in pKECY98 |
| pKECY103 | araC Para cra (T7gene10-RBS) in pBAD30 | cra (PCR OB161/OB162) in pKECY98 |
a
Features include Cmr (chloramphenicol resistance), Kanr (kanamycin resistance), MCS (multiple-cloning site), and pSC-repts (temperature-sensitive replication, derivative of pSC101).
b
Cloning was verified by sequencing of the cloned PCR fragments.