Basic Vector Information
- Vector Name:
- pEMG
- Antibiotic Resistance:
- Kanamycin
- Length:
- 3168 bp
- Type:
- Gene deletion vector
- Replication origin:
- R6K γ ori
- Source/Author:
- Martinez-Garcia E, de Lorenzo V.
pEMG 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 ℃.
pEMG vector Sequence
LOCUS 40924_17384 3168 bp DNA circular SYN 17-DEC-2018
DEFINITION Gene deletion vector pEMG, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3168)
AUTHORS Martinez-Garcia E, de Lorenzo V.
TITLE Engineering multiple genomic deletions in Gram-negative bacteria:
analysis of the multi-resistant antibiotic profile of Pseudomonas
putida KT2440
JOURNAL Environ. Microbiol. 13 (10), 2702-2716 (2011)
PUBMED 21883790
REFERENCE 2 (bases 1 to 3168)
AUTHORS Martinez-Garcia E, de Lorenzo V.
TITLE Direct Submission
JOURNAL Submitted (16-MAY-2011) Systems Biology Program, Centro Nacional de
Biotecnologia, C/ Darwin 3, Madrid 28049, Spain
REFERENCE 3 (bases 1 to 3168)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3168)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Environ.
Microbiol."; date: "2011"; volume: "13"; issue: "10"; pages:
"2702-2716"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-MAY-2011) Systems Biology Program, Centro Nacional de
Biotecnologia, C/ Darwin 3, Madrid 28049, Spain"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3168
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(126..917)
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
rep_origin complement(1281..1669)
/direction=LEFT
/label=R6K gamma ori
/note="gamma replication origin from E. coli plasmid R6K;
requires the R6K initiator protein pi for replication"
oriT 1829..1938
/label=oriT
/note="incP origin of transfer"
CDS 1971..2339
/codon_start=1
/label=traJ
/note="oriT-recognizing protein"
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
primer_bind 2774..2790
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 2792..2809
/label=I-SceI site
/note="I-SceI site"
misc_feature 2812..2868
/label=MCS
/note="pUC18/19 multiple cloning site"
misc_feature 2870..2887
/label=I-SceI site
/note="I-SceI site"
primer_bind complement(2902..2918)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(2926..2942)
/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)."
promoter complement(2950..2980)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(2995..3016)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
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