Basic Vector Information
- Vector Name:
- pOSIP-KO
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6472 bp
- Type:
- Cloning Vectors
- Replication origin:
- R6K γ ori
- Source/Author:
- St-Pierre F, Cui L, Priest DG, Endy D, Dodd IB, Shearwin KE.
- Copy Number:
- High copy number
pOSIP-KO vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
pOSIP-KO vector Sequence
LOCUS 40924_33912 6472 bp DNA circular SYN 18-DEC-2018
DEFINITION Chromosomal integration vector pOSIP-KO, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6472)
AUTHORS St-Pierre F, Cui L, Priest DG, Endy D, Dodd IB, Shearwin KE.
TITLE One-step cloning and chromosomal integration of DNA
JOURNAL ACS Synth Biol 2 (9), 537-541 (2013)
PUBMED 24050148
REFERENCE 2 (bases 1 to 6472)
AUTHORS St-Pierre F, Cui L, Priest DG, Endy D, Dodd IB, Shearwin KE.
TITLE Direct Submission
JOURNAL Submitted (12-MAY-2013) Molecular and Biomedical Science, The
University of Adelaide, Molecular Life Sciences Building, Adelaide,
South Australia 5005, Australia
REFERENCE 3 (bases 1 to 6472)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6472)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "ACS Synth
Biol"; date: "2013"; volume: "2"; issue: "9"; pages: "537-541"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-MAY-2013) Molecular and Biomedical Science, The University of
Adelaide, Molecular Life Sciences Building, Adelaide, South
Australia 5005, Australia"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6472
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 142..444
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
rep_origin 477..1065
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature 1125..1145
/label=BioBrick suffix
/note="universal suffix for all parts"
terminator 1200..1227
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(1276..1307)
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
protein_bind 1384..1417
/label=FRT (minimal)
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
CDS complement(1427..2137)
/label=lambda repressor (ts)
/note="temperature-sensitive variant of the phage lambda
repressor"
RBS 2226..2234
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
RBS 2246..2268
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2276..3283
/label=phage 186 integrase
/note="integrase from phage 186"
CDS 3460..4272
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 4397..4785
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
misc_feature 4796..4843
/note="FRT; Flp recombinase target site"
protein_bind 4796..4843
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
protein_bind complement(4856..5070)
/label=phage 186 attP
/note="attachment site of phage 186"
terminator complement(5187..5433)
/label=lambda tL3 terminator
/note="transcription terminator tL3 from phage lambda"
terminator 5753..5839
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 5934..6020
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 6115..6201
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator complement(6299..6342)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
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