Basic Vector Information
- Vector Name:
- pPK115
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6372 bp
- Type:
- Expression vector
- Replication origin:
- ori
- Source/Author:
- Coy DL, Hancock WO, Wagenbach M, Howard J.
pPK115 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 ℃.
pPK115 vector Sequence
LOCUS 40924_34729 6372 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pPK115, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6372)
AUTHORS Coy DL, Hancock WO, Wagenbach M, Howard J.
TITLE Kinesin's tail domain is an inhibitory regulator of the motor domain
JOURNAL Nat. Cell Biol. 1 (5), 288-292 (1999)
PUBMED 10559941
REFERENCE 2 (bases 1 to 6372)
AUTHORS Coy DL, Wagenbach M, Howard J.
TITLE Direct Submission
JOURNAL Submitted (30-DEC-1998) Department of Physiology and Biophysics,
University of Washington, Campus Box 357290, Seattle, WA 98195-7290,
USA
REFERENCE 3 (bases 1 to 6372)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6372)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Cell
Biol."; date: "1999"; volume: "1"; issue: "5"; pages: "288-292"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-DEC-1998) Department of Physiology and Biophysics, University of
Washington, Campus Box 357290, Seattle, WA 98195-7290, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6372
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 12..467
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 494..598
/label=AmpR promoter
CDS 599..1456
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1630..2218
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(2404..2546)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2651..2839)
/codon_start=1
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
/translation="VTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
promoter 3348..3366
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 3398..3420
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
RBS 4939..4947
/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 6158..6175
/codon_start=1
/label=thrombin site
/note="thrombin recognition and cleavage site"
/translation="LVPRGS"
CDS 6176..6193
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
CDS 6216..6233
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
terminator 6300..6347
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
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