pJP5603 vector (V005292)

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 Vector Map

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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

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Download GeneBank File(.gb)

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"