pCP20 vector map and sequence - novoprolabs.com
pCP20 is an ampicillin and CmR plasmid that shows temperature-sensitive replication and thermal induction of FLP synthesis. CmR and KmR mutants were transformed with pCP20, and ampicillin-resistant transformants were selected at 30°C, after which a few were colony-purified once nonselectively at 43°C and then tested for loss of all antibiotic ...
One-step inactivation of chromosomal genes in Escherichia coli …
pCP20 is an ampicillin and Cm R plasmid that shows temperature-sensitive replication and thermal induction of FLP synthesis . Cm R and Km R mutants were transformed with pCP20, and ampicillin-resistant transformants were selected at 30°C, after which a few were colony-purified once nonselectively at 43°C and then tested for loss of all ...
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Producing Gene Deletions in Escherichia coli by P1 Transduction …
The plasmid pCP20, encoding the FLP recombinase, was introduced into these clones and, subsequently, cured by growing at 43 °C in the absence of an antibiotic selection. A number of clones were screened for sensitivity to both Amp (which is a marker of pCP20) and Kan, and several clones sensitive to both were obtained.
ProcedureFLPFRTRecombination - Barrick Lab
Transform with pCP20. This plasmid has a temperature-sensitive origin of replication, confers ampicillin resistance, and encodes the FLP recombinase. Add 500 µl SOC and incubate in a 1.7 ml tube shaking on its side at ≥120 rpm for 1 hr at 30°C to allow antibiotic resistance induction.
Review of knockout technology approaches in bacterial drug …
Aug 23, 2023 · The resistance gene can be removed with the help of pCP20 plasmid, which expresses FLP recombinase that recognizes direct repeat FRT sites flanking the resistance gene (as shown in Fig. 1, adapted from (Datsenko & Wanner, 2000)). The temperature-sensitive replicons present in pKD46 and pCP20 plasmids can be eliminated by incubating them at 37 °C.
pCP20 - СAGER
pCP20. A plasmid for introducing deletions in E. coli genome. The plasmid carries gene coding for temperature-sensitive version of Rep101 protein needed for replication with the pSC101 origin.
pBAD-Flp - Addgene
Plasmid pBAD-Flp from Dr. Lydia Freddolino's lab contains the insert Flp recombinase and is published in Cell Syst. 2019 Mar 27;8(3):212-225.e9. doi: 10.1016/j.cels.2019.02.004. Epub 2019 Mar 20. This plasmid is available through Addgene.
CRISPR-FRT targets shared sites in a knock-out collection for off …
Jun 8, 2018 · A new pTarget plasmid encoding an sgRNA targeting a unique sequence on the recA plasmid could be transformed into the chromosomal recA library pool and plated on chloramphenicol + spectinomycin...
SlyD-deficient Escherichia coli strains: A highway to …
May 23, 2018 · pCP20 is an Ap R and Cm R plasmid that exhibits temperature-sensitive replication and thermal induction of FLP synthesis [14]. Km R mutants were transformed using pCP20, and ampicillin-resistant transformants were selected at 30 °C.