Basic Vector Information
- Vector Name:
- pOSIP-KP
- Antibiotic Resistance:
- Kanamycin
- Length:
- 6938 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-KP 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-KP vector Sequence
LOCUS 40924_33917 6938 bp DNA circular SYN 18-DEC-2018 DEFINITION Chromosomal integration vector pOSIP-KP, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6938) 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 6938) 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 6938) TITLE Direct Submission REFERENCE 4 (bases 1 to 6938) 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..6938 /mol_type="other DNA" /organism="synthetic DNA construct" CDS 151..453 /label=ccdB /note="CcdB, a bacterial toxin that poisons DNA gyrase" rep_origin 486..1074 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" misc_feature 1134..1154 /label=BioBrick suffix /note="universal suffix for all parts" terminator 1203..1230 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator complement(1279..1310) /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" protein_bind 1387..1420 /label=FRT (minimal) /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" CDS complement(1430..2140) /label=lambda repressor (ts) /note="temperature-sensitive variant of the phage lambda repressor" RBS 2229..2237 /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)" CDS 2372..3577 /label=phage phi-80 integrase /note="integrase from phage phi-80" CDS 3757..4569 /label=KanR /note="aminoglycoside phosphotransferase" rep_origin 4694..5082 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" protein_bind 5093..5140 /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)." misc_feature 5107..5140 /note="FRT; Flp recombinase target site" protein_bind complement(5247..5564) /label=phage Phi-80 attP /note="attachment site of phage phi-80" terminator complement(5653..5899) /label=lambda tL3 terminator /note="transcription terminator tL3 from phage lambda" terminator 6219..6305 /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 6400..6486 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 6581..6667 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator complement(6765..6808) /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene"
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