pEMG vector (V007569)

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 Map

pEMG3168 bp6001200180024003000NeoR/KanRR6K gamma orioriTtraJM13 fwdI-SceI siteMCSI-SceI siteM13 revlac operatorlac promoterCAP binding site

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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