Basic Vector Information
- Vector Name:
- p283
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5282 bp
- Type:
- Structural Genomics Vectors
- Replication origin:
- ori
- Source/Author:
- Cormier CY, Mohr SE, Zuo D, Hu Y, Rolfs A, Kramer J, Taycher E,
- Copy Number:
- High copy number
p283 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 ℃.
p283 vector Sequence
LOCUS p283. 5282 bp DNA circular SYN 01-JAN-1980
DEFINITION Bacterial vector for expressing a protein with an C-terminal
TEV-7xHis cassette.
ACCESSION .
VERSION .
KEYWORDS p283.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5282)
AUTHORS Cormier CY, Mohr SE, Zuo D, Hu Y, Rolfs A, Kramer J, Taycher E,
Kelley F, Fiacco M, Turnbull G, LaBaer J.
TITLE Protein Structure Initiative Material Repository: an open shared
public resource of structural genomics plasmids for the biological
community.
JOURNAL Nucleic Acids Res. 2010;38:D743-9.
PUBMED 19906724
REFERENCE 2 (bases 1 to 5282)
AUTHORS Midwest Center for Structural Genomics
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5282)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic
Acids Res."; date: "2010"; volume: "38"; pages: "D743-9"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5282
/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"
CDS complement(563..1375)
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 1497..2085
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(2271..2413)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2518..2706)
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
protein_bind complement(3481..3502)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(3518..4597)
/label=lacI
/note="lac repressor"
promoter complement(4598..4675)
/label=lacI promoter
promoter 4984..5002
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5003..5027
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
RBS 5042..5064
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5072..5074
/codon_start=1
/product="start codon"
/label=start codon
/note="ATG"
/translation="M"
misc_feature 5075..5092
/label=MCS
/note="MCS"
/note="multiple cloning site"
CDS 5096..5116
/label=TEV site
/note="tobacco etch virus (TEV) protease recognition and
cleavage site"
CDS 5123..5143
/label=7xHis
/note="6xHis affinity tag"
terminator 5210..5257
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
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