LOCUS       40924_14865        6820 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pDMK3, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6820)
  AUTHORS   Kadzhaev K, Zingmark C, Golovliov I, Bolanowski M, Shen H, Conlan W,
            Sjostedt A.
  TITLE     Identification of genes contributing to the virulence of Francisella
            tularensis SCHU S4 in a mouse intradermal infection model
  JOURNAL   PLoS ONE 4 (5), E5463 (2009)
  PUBMED    19424499
REFERENCE   2  (bases 1 to 6820)
  AUTHORS   Kadzhaev K, Golovliov I, Sjostedt A.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAR-2009) Clinical Bacteriology, Umea University, NUS,
            Umea 90185, Sweden
REFERENCE   3  (bases 1 to 6820)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6820)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2009"; volume: "4"; issue: "5"; pages: "E5463"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (13-MAR-2009) Clinical Bacteriology, Umea University, NUS, Umea 
            90185, Sweden"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6820
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(120..136)
                     /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)."
     rep_origin      complement(171..559)
                     /direction=LEFT
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K;
                     requires the R6K initiator protein pi for replication"
     CDS             complement(585..1142)
                     /codon_start=1
                     /gene="traI"
                     /product="TraI"
                     /label=traI
                     /protein_id="ACO50702.1"
                     /translation="MIAKHVPMRSIKKSDFAELVKYITDEQGKTERLGHVRVTNCEANT
                     LPAVMAEVMATQHGNTRSEADKTYHLLVSFRAGEKPDAETLRAIEDRICAGLGFAEHQR
                     VSAVHHDTDNLHIHIAINKIHPTRNTIHEPYRAYRALADLCATLERDYGLERDNHETRQ
                     RVSENRANDMERHAGVESLVGWI"
     gene            complement(585..1142)
                     /gene="traI"
                     /label=traI
     CDS             complement(1139..1180)
                     /codon_start=1
                     /gene="traX"
                     /product="TraX"
                     /label=traX
                     /protein_id="ACO50703.1"
                     /translation="MTFLAAKTAGGCA"
     gene            complement(1139..1180)
                     /gene="traX"
                     /label=traX
     CDS             complement(1180..1548)
                     /label=traJ
                     /note="oriT-recognizing protein"
     oriT            complement(1581..1690)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             1916..2312
                     /codon_start=1
                     /gene="traK"
                     /product="TraK"
                     /label=traK
                     /protein_id="ACO50705.1"
                     /translation="MPKSYTDELAEWVESRAAKKRRRDEAAVAFLAVRADVEAALASGY
                     ALVTIWEHMRETGKVKFSYETFRSHARRHIKAKPADVPAPQAKAAEPAPAPKTPEPRRP
                     KQGGKAEKPAPAAAPTGFTFNPTPDKKD"
     gene            1916..2312
                     /gene="traK"
                     /label=traK
     promoter        3127..3572
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             3573..4991
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
     regulatory      5711..6018
                     /label=groES promoter
                     /note="groES promoter"
                     /regulatory_class="promoter"
     CDS             6026..6817
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
ORIGIN
        1 attgtcgacc ctcgagtacg cgtcactagt ggggcccttc tagatagatc ttgcatgcgg
       61 ccgcatgcat cgcgagtact gcggccgcgt aacctgagct ctcccgggaa ttccacaaat
      121 tgttatccgc tcacaattcc acatgtggaa ttccacatgt ggaattccca tgtcagccgt
      181 taagtgttcc tgtgtcactc aaaattgctt tgagaggctc taagggcttc tcagtgcgtt
      241 acatccctgg cttgttgtcc acaaccgtta aaccttaaaa gctttaaaag ccttatatat
      301 tctttttttt cttataaaac ttaaaacctt agaggctatt taagttgctg atttatatta
      361 attttattgt tcaaacatga gagcttagta cgtgaaacat gagagcttag tacgttagcc
      421 atgagagctt agtacgttag ccatgagggt ttagttcgtt aaacatgaga gcttagtacg
      481 ttaaacatga gagcttagta cgtgaaacat gagagcttag tacgtactat caacaggttg
      541 aactgctgat cttcagatcc tctacgccgg acgcatcgtg gccggatcca gccgaccagg
      601 ctttccacgc ccgcgtgccg ctccatgtcg ttcgcgcggt tctcggaaac gcgctgccgc
      661 gtttcgtgat tgtcacgctc aagcccgtag tcccgttcga gcgtcgcgca gaggtcagcg
      721 agggcgcggt aggcccgata cggctcatgg atggtgtttc gggtcgggtg aatcttgttg
      781 atggcgatat ggatgtgcag gttgtcggtg tcgtgatgca cggcactgac gcgctgatgc
      841 tcggcgaagc caagcccagc gcagatgcgg tcctcaatcg cgcgcaacgt ctccgcgtcg
      901 ggcttctctc ccgcgcggaa gctaaccagc aggtgatagg tcttgtcggc ctcggaacgg
      961 gtgttgccgt gctgggtcgc catcacctcg gccatgacag cgggcagggt gtttgcctcg
     1021 cagttcgtga cgcgcacgtg acccaggcgc tcggtcttgc cttgctcgtc ggtgatgtac
     1081 ttcaccagct ccgcgaagtc gctcttcttg atggagcgca tggggacgtg cttggcaatc
     1141 acgcgcaccc cccggccgtt ttagcggcta aaaaagtcat ggctctgccc tcgggcggac
     1201 cacgcccatc atgaccttgc caagctcgtc ctgcttctct tcgatcttcg ccagcagggc
     1261 gaggatcgtg gcatcaccga accgcgccgt gcgcgggtcg tcggtgagcc agagtttcag
     1321 caggccgccc aggcggccca ggtcgccatt gatgcgggcc agctcgcgga cgtgctcata
     1381 gtccacgacg cccgtgattt tgtagccctg gccgacggcc agcaggtagg ccgacaggct
     1441 catgccggcc gccgccgcct tttcctcaat cgctcttcgt tcgtctggaa ggcagtacac
     1501 cttgataggt gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc
     1561 atctgttacg ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc
     1621 caggtgcgaa taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc
     1681 tatcctgccc ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca
     1741 aaatcctgta tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc
     1801 gaagcagggt tatgcagcgg aaaagcgctg cttccctgct gttttgtgga atatctaccg
     1861 actggaaaca ggcaaatgca ggaaattact gaactgaggg gacaggcgag agacgatgcc
     1921 aaagagctac accgacgagc tggccgagtg ggttgaatcc cgcgcggcca agaagcgccg
     1981 gcgtgatgag gctgcggttg cgttcctggc ggtgagggcg gatgtcgagg cggcgttagc
     2041 gtccggctat gcgctcgtca ccatttggga gcacatgcgg gaaacgggga aggtcaagtt
     2101 ctcctacgag acgttccgct cgcacgccag gcggcacatc aaggccaagc ccgccgatgt
     2161 gcccgcaccg caggccaagg ctgcggaacc cgcgccggca cccaagacgc cggagccacg
     2221 gcggccgaag caggggggca aggctgaaaa gccggccccc gctgcggccc cgaccggctt
     2281 caccttcaac ccaacaccgg acaaaaagga tcctctacgc cggacgcatc gtggccggca
     2341 tcaccggcgc cacaggtgcg gttgctggcg cctatatcgc cgacatcacc gatggggaag
     2401 atcgggctcg ccacttcggg ctcatgagcg cttgtttcgg cgtgggtatg gtggcaggcc
     2461 ccgtggccgg gggactgttg ggcgccatct ccttgctgcc tcgcgcgttt cggtgatgac
     2521 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     2581 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggcgca
     2641 gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat gcggcatcag
     2701 agcagattgt actgagagtg caccataaat caatctaaag tatatatgag taaacttggt
     2761 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2821 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2881 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     2941 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3001 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3061 tgcgcaacgt tgttgccatt gctgcagcca agctagacct aggccttaag atccttttta
     3121 acccatcaca tatacctgcc gttcactatt atttagtgaa atgagatatt atgatatttt
     3181 ctgaattgtg attaaaaagg caactttatg cccatgcaac agaaactata aaaaatacag
     3241 agaatgaaaa gaaacagata gattttttag ttctttaggc ccgtagtctg caaatccttt
     3301 tatgattttc tatcaaacaa aagaggaaaa tagaccagtt gcaatccaaa cgagagtcta
     3361 atagaatgag gtcgaaaagt aaatcgcgcg ggtttgttac tgataaagca ggcaagacct
     3421 aaaatgtgta aagggcaaag tgtatacttt ggcgtcaccc cttacatatt ttaggtcttt
     3481 ttttattgtg cgtaactaac ttgccatctt caaacaggag ggctggaaga agcagaccgc
     3541 taacacagta cataaaaaag gagacatgaa cgatgaacat caaaaagttt gcaaaacaag
     3601 caacagtatt aacctttact accgcactgc tggcaggagg cgcaactcaa gcgtttgcga
     3661 aagaaacgaa ccaaaagcca tataaggaaa catacggcat ttcccatatt acacgccatg
     3721 atatgctgca aatccctgaa cagcaaaaaa atgaaaaata tcaagttcct gaattcgatt
     3781 cgtccacaat taaaaatatc tcttctgcaa aaggcctgga cgtttgggac agctggccat
     3841 tacaaaacgc tgacggcact gtcgcaaact atcacggcta ccacatcgtc tttgcattag
     3901 ccggagatcc taaaaatgcg gatgacacat cgatttacat gttctatcaa aaagtcggcg
     3961 aaacttctat tgacagctgg aaaaacgctg gccgcgtctt taaagacagc gacaaattcg
     4021 atgcaaatga ttctatccta aaagaccaaa cacaagaatg gtcaggttca gccacattta
     4081 catctgacgg aaaaatccgt ttattctaca ctgatttctc cggtaaacat tacggcaaac
     4141 aaacactgac aactgcacaa gttaacgtat cagcatcaga cagctctttg aacatcaacg
     4201 gtgtagagga ttataaatca atctttgacg gtgacggaaa aacgtatcaa aatgtacagc
     4261 agttcatcga tgaaggcaac tacagctcag gcgacaacca tacgctgaga gatcctcact
     4321 acgtagaaga taaaggccac aaatacttag tatttgaagc aaacactgga actgaagatg
     4381 gctaccaagg cgaagaatct ttatttaaca aagcatacta tggcaaaagc acatcattct
     4441 tccgtcaaga aagtcaaaaa cttctgcaaa gcgataaaaa acgcacggct gagttagcaa
     4501 acggcgctct cggtatgatt gagctaaacg atgattacac actgaaaaaa gtgatgaaac
     4561 cgctgattgc atctaacaca gtaacagatg aaattgaacg cgcgaacgtc tttaaaatga
     4621 acggcaaatg gtacctgttc actgactccc gcggatcaaa aatgacgatt gacggcatta
     4681 cgtctaacga tatttacatg cttggttatg tttctaattc tttaactggc ccatacaagc
     4741 cgctgaacaa aactggcctt gtgttaaaaa tggatcttga tcctaacgat gtaaccttta
     4801 cttactcaca cttcgctgta cctcaagcga aaggaaacaa tgtcgtgatt acaagctata
     4861 tgacaaacag aggattctac gcagacaaac aatcaacgtt tgcgccaagc ttcctgctga
     4921 acatcaaagg caagaaaaca tctgttgtca aagacagcat ccttgaacaa ggacaattaa
     4981 cagttaacaa ataaaaacgc aaaagaaaat gccgatatcc tattggcatt ttcttttatt
     5041 tcttatcaac ataaaggtga atcccatatg aactatataa aagcaggcaa atggctaacc
     5101 gtattcctaa ccttttggta atgactccaa cttattgata gtgttttatg ttcagataat
     5161 gcccgatgac tttgtcatgc agctccaccg attttgagaa cgacagcgac ttccgtccca
     5221 gccgtgccag gtgctgcctc agattcaggt tatgccgctc aattcgctgc gtatatcgct
     5281 tgctgattac gtgcagcttt cccttcaggc gggattcata cagcggccag ccatccgtca
     5341 tccatatcac cacgtcaaag ggtgacagca ggctcataag acgccccagc gtcgccatag
     5401 tgcgttcacc gaatacgtgc gcaacaaccg tcttccggag actgtcatac gcgtaaaaca
     5461 gccagcgctg gcgcgattta gccccgacat agccccactg ttcgtccatt tccgcgcaga
     5521 cgatgacgtc actgcccggc tgtatgcgcg aggttaccga ctgcggcctg agttttttaa
     5581 gtgacgtaaa atcgtgttga ggccaacgcc cataatgcgg gctgttgccc ggcatccaac
     5641 gccattcatg gccatatcaa tgattttctg gtgcgtaccg ggttgagaag cggtgtaagt
     5701 gaactgcagg ccaaaaaact caaatcctat tgtatggatt agtcgagcta aaaagctcat
     5761 attttttata ttcaaactat atacccttat ttacaagctt tgaaaaataa acttaattat
     5821 tatatatgtt atttagctag tttttttaat taaagttaaa atcgagagct tgtttgacaa
     5881 aaaaacaaaa aaatttcttg aaaatttttt tttgactcaa tatctagact tgcaagagct
     5941 tggagctttg agattgttct aagatgcaca caaattcaaa acgcttaaac aaaaaactta
     6001 atttaacaaa ggagtaagag gatccatgat tgaacaagat ggattgcacg caggttctcc
     6061 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     6121 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     6181 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac
     6241 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     6301 gctattgggc gaagtgccgg ggcaggatct cctgtcatcc caccttgctc ctgccgagaa
     6361 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     6421 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     6481 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     6541 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     6601 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     6661 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     6721 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     6781 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
//
