Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Orr Schlesinger
Date created: 2013-09-09 11:00:00
Date modified: 2015-05-08 01:09:43
pKD46 functional unit
| Types | DnaRegion |
| Roles | Device engineered_region |
| Sequences | BBa_K1223012_sequence (Version 1) |
Description
pKD46 functional part - the plasmid backbone without antibiotic resistance.pKD plasmids are carrying the Lambda Red recombineering system. this system is highly efficient in making indels in E.coli genome.
just need to assemble with selection gene of choice to get any plasmid from the pKD family - pKD46 carries the bla gene (ampicillin resistance), pKD78 carries the cat gene(chloramphenicol resistance), pKD119 carries the tet gene (tetracycline resistance)or create new plasmid with kanamicin resistance.
the part includes:
- AraC repressor CDS.
- beta gamma and exo genes of the lambda bacteriophage regulated by AraBAD promoter (arabinose induction).
- OriR101 replication origin - temperature sensitive origin. grow the bacteria at 30C to keep and amplify the plasmid or grow at 42C to eliminate it.
Kirill A. Datsenko and Barry L. Wanner
One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products
PNAS 2000 97 (12) 6640-6645; published ahead of print May 30, 2000, doi:10.1073/pnas.120163297
Notes
just need to assemble with selection gene of choice to get any plasmid from the pKD family - pKD46 carries the bla gene (ampicillin resistance), pKD78 carries the cat gene(chloramphenicol resistance), pKD119 carries the tet gene (tetracycline resistance)or create new plasmid with kanamicin resistance.temperature sensitive origin of replication. grow the bacteria at 30C to keep and amplify the plasmid or grow at 42C to eliminate it.
Source
genbank.Kirill A. Datsenko and Barry L. Wanner
One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products
PNAS 2000 97 (12) 6640-6645; published ahead of print May 30, 2000, doi:10.1073/pnas.120163297
| Sequence Annotation | Location | Component / Role(s) |
| AraC CDS gamma protein CDS beta protein CDS exonuclease CDS RepA101 protein CDS OriR101 replication origin | 3,881 1236,1652 1658,2443 2440,3120 3488,4438 4442,5182 | feature/cds CDS CDS feature/cds CDS feature/cds feature/cds CDS feature/cds CDS feature/misc sequence_feature |