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."
ORIGIN
        1 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
       61 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc
      121 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
      181 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
      241 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
      301 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
      361 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
      421 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
      481 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
      541 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
      601 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
      661 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
      721 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
      781 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
      841 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
      901 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
      961 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     1021 accttaccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc
     1081 cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg
     1141 ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc
     1201 ggactggctt tctacgtgtt ccgcttcctt tagcagccct tgcgccctga gtgcttgcgg
     1261 cagcgtgaag ctaattccca tgtcagccgt taagtgttcc tgtgtcactc aaaattgctt
     1321 tgagaggctc taagggcttc tcagtgcgtt acatccctgg cttgttgtcc acaaccgtta
     1381 aaccttaaaa gctttaaaag ccttatatat tctttttttt cttataaaac ttaaaacctt
     1441 agaggctatt taagttgctg atttatatta attttattgt tcaaacatga gagcttagta
     1501 cgtgaaacat gagagcttag tacgttagcc atgagagctt agtacgttag ccatgagggt
     1561 ttagttcgtt aaacatgaga gcttagtacg ttaaacatga gagcttagta cgtgaaacat
     1621 gagagcttag tacgtactat caacaggttg aactgctgat cttcagatcc tctacgccgg
     1681 acgcatcgtg gccgttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt
     1741 cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact
     1801 ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga
     1861 gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg
     1921 ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg atggctgatg
     1981 aaaccaagcc aaccaggaag ggcagcccac ctatcaaggt gtactgcctt ccagacgaac
     2041 gaagagcgat tgaggaaaag gcggcggcgg ccggcatgag cctgtcggcc tacctgctgg
     2101 ccgtcggcca gggctacaaa atcacgggcg tcgtggacta tgagcacgtc cgcgagctgg
     2161 cccgcatcaa tggcgacctg ggccgcctgg gcggcctgct gaaactctgg ctcaccgacg
     2221 acccgcgcac ggcgcggttc ggtgatgcca cgatcctcgc cctgctggcg aagatcgaag
     2281 agaagcagga cgagcttggc aaggtcatga tgggcgtggt ccgcccgagg gcagagccat
     2341 gactttttta gccgctaaaa cggccggggg gtgcgcgtga ttgccaagca cgtccccatg
     2401 cgctccatca agaagagcga cttcgcggag ctggtgaagt acatcaccga cgagcaaggc
     2461 aagaccgacc aaagcggcca tcgtgcctcc ccactcctgc agttcggggg catggatgcg
     2521 cggatagccg ctgctggttt cctggatgcc gacggatttg cactgccggt agaactccgc
     2581 gaggtcgtcc agcctcaggc agcagctgaa ccaactcgcg aggggatcga gccccattcg
     2641 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     2701 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     2761 cagtcacgac gttgtaaaac gacggccagt atagggataa cagggtaatc tgaattcgag
     2821 ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttct agggataaca
     2881 gggtaatccg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     2941 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     3001 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     3061 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattggggg
     3121 gtgggcgaag aactccagca tgagatcccc gcgctggagg atcatcca
//
