Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V005292 | pJP5603 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pJP5603
- Antibiotic Resistance:
- Kanamycin
- Length:
- 3126 bp
- Type:
- Suicide vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Riedel T, Rohlfs M, Buchholz I, Wagner-Dobler I, Reck M.
- Promoter:
- lac
pJP5603 vector Map
Plasmid 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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
pJP5603 vector Sequence
LOCUS 40924_26326 3126 bp DNA circular SYN 18-DEC-2018
DEFINITION Suicide vector pJP5603, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3126)
AUTHORS Riedel T, Rohlfs M, Buchholz I, Wagner-Dobler I, Reck M.
TITLE Complete sequence of the suicide vector pJP5603
JOURNAL Plasmid 69 (1), 104-107 (2013)
PUBMED 22902299
REFERENCE 2 (bases 1 to 3126)
AUTHORS Riedel T, Reck M.
TITLE Direct Submission
JOURNAL Submitted (12-MAR-2012) Microbial Communication, Helmholtz-Centre
for Infection Research, Inhoffenstrasse 7, Braunschweig, Lower
Saxony 38124, Germany
REFERENCE 3 (bases 1 to 3126)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3126)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid";
date: "2013"; volume: "69"; issue: "1"; pages: "104-107"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(12-MAR-2012) Microbial Communication, Helmholtz-Centre for
Infection Research, Inhoffenstrasse 7, Braunschweig, Lower Saxony
38124, Germany"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3126
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS 204..995
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
protein_bind 1273..1294
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 1309..1339
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1347..1363
/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)."
primer_bind 1371..1387
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(1400..1456)
/label=MCS
/note="pUC18/19 multiple cloning site"
primer_bind complement(1457..1473)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(1908..2276)
/codon_start=1
/label=traJ
/note="oriT-recognizing protein"
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
oriT complement(2309..2418)
/direction=LEFT
/label=oriT
/note="incP origin of transfer"
rep_origin 2578..2966
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"