LOCUS       Exported                3168 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Gene deletion vector pEMG, complete sequence.
ACCESSION   JF965437
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3168
                     /organism="Gene deletion vector pEMG"
                     /mol_type="other DNA"
                     /db_xref="taxon:1042310"
     CDS             complement(123..917)
                     /codon_start=1
                     /gene="nptII"
                     /product="aminoglycoside 3-phosphotransferase"
                     /label=nptII
                     /note="kanamycin resistance protein"
                     /protein_id="AEH59866.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(123..917)
                     /gene="nptII"
                     /label=nptII
     rep_origin      1278..1669
                     /label=R6KoriV
                     /note="R6KoriV"
     rep_origin      1281..1669
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     misc_feature    1695..1937
                     /note="oriT; origin of transference"
     oriT            1829..1938
                     /note="incP origin of transfer"
     CDS             1971..2342
                     /codon_start=1
                     /gene="traJ"
                     /product="conjugal transfer relaxosome component"
                     /label=traJ
                     /note="TraJ"
                     /protein_id="AEH59867.1"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     gene            1971..2342
                     /gene="traJ"
                     /label=traJ
     CDS             complement(2595..2906)
                     /codon_start=1
                     /gene="lacZ alpha"
                     /product="LacZ alpha peptide"
                     /label=lacZ alpha
                     /note="has I-SceI sites"
                     /protein_id="AEH59868.1"
                     /translation="MTMITPDYPVIPRSLHACRSTLEDPRVPSSNSDYPVIPILAVVLQ
                     RRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWGSIPSRVGSAAA"
     gene            complement(2595..2906)
                     /gene="lacZ alpha"
                     /label=lacZ alpha
     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    2810..2869
                     /label=multiple cloning site (MCS)
                     /note="multiple cloning site (MCS)"
     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    2926..2942
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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    2995..3016
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."