Basic Vector Information
- Vector Name:
- pNT562
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5058 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Nishida T, Watanabe K, Tachibana M, Shimizu T, Watarai M.
- Promoter:
- tac
pNT562 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 ℃.
pNT562 vector Sequence
LOCUS 40924_33547 5058 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pNT562 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5058)
AUTHORS Nishida T, Watanabe K, Tachibana M, Shimizu T, Watarai M.
TITLE Characterization of the cryptic plasmid pOfk55 from Legionella
pneumophila and construction of a pOfk55-derived shuttle vector
JOURNAL Plasmid 90, 30-37 (2017)
PUBMED 28259635
REFERENCE 2 (bases 1 to 5058)
AUTHORS Nishida T, Watanabe K, Tachibana M, Shimizu T, Watarai M.
TITLE Direct Submission
JOURNAL Submitted (06-FEB-2017) Contact:Takashi Nishida Yamaguchi
University, The United Graduate School of Veterinary Science; 1677-1
Yoshida, Yamaguchi, Yamaguchi 753-8515, Japan
REFERENCE 3 (bases 1 to 5058)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5058)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 90,
30-37 (2017)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(06-FEB-2017) Contact:Takashi Nishida Yamaguchi University, The
United Graduate School of Veterinary Science; 1677-1 Yoshida,
Yamaguchi, Yamaguchi 753-8515, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5058
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 563..572
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1016..1603
/codon_start=1
/product="putative replication protein"
/label=putative replication protein
/protein_id="BAX01899.1"
/translation="MKKEPIPGKRKNVLAYPENPFWQKTEIKIGSKMVKVSGGKHINIE
GESISHSGIHVVKEIDENEFVKIYTKNIKAIFDLKPTAQRVLQYLITELQKTPNADAVY
LAWVGAEEYFSENHIKSSRASFHRALSELIQKGFLAESTKPNMFWFNPNLFFNGNRLTF
IHEYRKKTIQEIGKEENLVQSDIDKQIQIIFN"
CDS complement(1678..2757)
/label=lacI
/note="lac repressor"
promoter complement(2758..2833)
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
promoter 3067..3095
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 3103..3119
/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)."
misc_feature complement(3166..3273)
/label=MCS
/note="pBluescript multiple cloning site"
CDS 3479..4270
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
rep_origin 4452..5040
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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