LOCUS       Exported                7107 bp DNA     circular SYN 10-JUL-2025
DEFINITION  Cloning vector pDM4, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7107)
  AUTHORS   Mou X, Spinard EJ, Driscoll MV, Zhao W, Nelson DR.
  TITLE     H-NS Is a Negative Regulator of the Two Hemolysin/Cytotoxin Gene 
            Clusters in Vibrio anguillarum
  JOURNAL   Infect. Immun. 81 (10), 3566-3576 (2013)
  PUBMED    23836825
REFERENCE   2  (bases 1 to 7107)
  AUTHORS   Mou X, Spinard EJ, Driscoll MV, Zhao W, Nelson DR.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAR-2013) Cell and Molecular Biology, University of 
            Rhode Island, 120 Flagg Rd, Kingston, RI 02881, USA
REFERENCE   3  (bases 1 to 7107)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7107)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 7107)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Infect.
            Immun."; date: "2013"; volume: "81"; issue: "10"; pages: "3566-3576"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (19-MAR-2013) Cell and Molecular Biology, University of Rhode
            Island, 120 Flagg Rd, Kingston, RI 02881, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            623..732
                     /label=oriT
                     /note="incP origin of transfer"
     CDS             765..1133
                     /codon_start=1
                     /label=traJ
                     /note="oriT-recognizing protein"
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             1192..1818
                     /codon_start=1
                     /product="conjugal transfer relaxase TraI"
                     /label=conjugal transfer relaxase TraI
                     /protein_id="AGO64135.1"
                     /translation="MRSIKKSDFAELVKYITDEQGKTERLGHVRVTNCEANTLPAVMAE
                     VMATQHGNTRSEADKTYHLLVSFRAGEKPDAETLRAIEDRICAGLGFAEHQRVSAVHHD
                     TDNLHIHIAINKIHPTRNTIHEPYRAYRALADLCATLERDYGLERDNHETRQRVSENRA
                     NDMERHAGVESLVGWIRPRCVRRRGSEDQQFNLLIVRTKLSCFTY"
     rep_origin      1754..2142
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    2177..2193
                     /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        2575..2677
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             2678..3334
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        3756..4201
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             4202..5620
                     /codon_start=1
                     /label=SacB
                     /note="secreted levansucrase that renders bacterial growth 
                     sensitive to sucrose"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     CDS             complement(5761..6138)
                     /codon_start=1
                     /product="insertion element IS1 1/5/6 protein insB"
                     /label=insertion element IS1 1/5/6 protein insB
                     /protein_id="AGO64138.1"
                     /translation="MDEQWGYVGAKSRQRWLFYAYDSLRKTVVAHVFGERTMATLGRLM
                     SLLSPFDVVIWMTDGWPLYESRLKGKLHVISKRYTQRIERHNLNLRQHLARLGRKSLSF
                     SKSVELHDKVIGHYLNIKHYQ"
     CDS             6416..6652
                     /codon_start=1
                     /product="beta-lactamase"
                     /EC_number="3.5.2.6"
                     /label=beta-lactamase
                     /protein_id="AGO64139.1"
                     /translation="MEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKP
                     SRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
ORIGIN
        1 gatccttttt gtccggtgtt gggttgaagg tgaagccggt cggggccgca gcgggggccg
       61 gcttttcagc cttgcccccc tgcttcggcc gccgtggctc cggcgtcttg ggtgccggcg
      121 cgggttccgc agccttggcc tgcggtgcgg gcacatcggc gggcttggcc ttgatgtgcc
      181 gcctggcgtg cgagcggaac gtctcgtagg agaacttgac cttccccgtt tcccgcatgt
      241 gctcccaaat ggtgacgagc gcatagccgg acgctaacgc cgcctcgaca tccgccctca
      301 ccgccaggaa cgcaaccgca gcctcatcac gccggcgctt cttggccgcg cgggattcaa
      361 cccactcggc cagctcgtcg gtgtagctct ttggcatcgt ctctcgcctg tcccctcagt
      421 tcagtaattt cctgcatttg cctgtttcca gtcggtagat attccacaaa acagcaggga
      481 agcagcgctt ttccgctgca taaccctgct tcggggtcat tatagcgatt ttttcggtat
      541 atccatcctt tttcgcacga tatacaggat tttgccaaag ggttcgtgta gactttcctt
      601 ggtgtatcca acggcgtcag ccgggcagga taggtgaagt aggcccaccc gcgagcgggt
      661 gttccttctt cactgtccct tattcgcacc tggcggtgct caacgggaat cctgctctgc
      721 gaggctggcc ggctaccgcc ggcgtaacag atgagggcaa gcggatggct gatgaaacca
      781 agccaaccag gaagggcagc ccacctatca aggtgtactg ccttccagac gaacgaagag
      841 cgattgagga aaaggcggcg gcggccggca tgagcctgtc ggcctacctg ctggccgtcg
      901 gccagggcta caaaatcacg ggcgtcgtgg actatgagca cgtccgcgag ctggcccgca
      961 tcaatggcga cctgggccgc ctgggcggcc tgctgaaact ctggctcacc gacgacccgc
     1021 gcacggcgcg gttcggtgat gccacgatcc tcgccctgct ggcgaagatc gaagagaagc
     1081 aggacgagct tggcaaggtc atgatgggcg tggtccgccc gagggcagag ccatgacttt
     1141 tttagccgct aaaacggccg gggggtgcgc gtgattgcca agcacgtccc catgcgctcc
     1201 atcaagaaga gcgacttcgc ggagctggtg aagtacatca ccgacgagca aggcaagacc
     1261 gagcgcctgg gtcacgtgcg cgtcacgaac tgcgaggcaa acaccctgcc cgctgtcatg
     1321 gccgaggtga tggcgaccca gcacggcaac acccgttccg aggccgacaa gacctatcac
     1381 ctgctggtta gcttccgcgc gggagagaag cccgacgcgg agacgttgcg cgcgattgag
     1441 gaccgcatct gcgctgggct tggcttcgcc gagcatcagc gcgtcagtgc cgtgcatcac
     1501 gacaccgaca acctgcacat ccatatcgcc atcaacaaga ttcacccgac ccgaaacacc
     1561 atccatgagc cgtatcgggc ctaccgcgcc ctcgctgacc tctgcgcgac gctcgaacgg
     1621 gactacgggc ttgagcgtga caatcacgaa acgcggcagc gcgtttccga gaaccgcgcg
     1681 aacgacatgg agcggcacgc gggcgtggaa agcctggtcg gctggatccg gccacgatgc
     1741 gtccggcgta gaggatctga agatcagcag ttcaacctgt tgatagtacg tactaagctc
     1801 tcatgtttca cgtactaagc tctcatgttt aacgtactaa gctctcatgt ttaacgaact
     1861 aaaccctcat ggctaacgta ctaagctctc atggctaacg tactaagctc tcatgtttca
     1921 cgtactaagc tctcatgttt gaacaataaa attaatataa atcagcaact taaatagcct
     1981 ctaaggtttt aagttttata agaaaaaaaa gaatatataa ggcttttaaa gcttttaagg
     2041 tttaacggtt gtggacaaca agccagggat gtaacgcact gagaagccct tagagcctct
     2101 caaagcaatt ttgagtgaca caggaacact taacggctga catgggaatt ccacatgtgg
     2161 aattccacat gtggaattgt gagcggataa caatttgtgg aattcccggg agagctcagg
     2221 ttacccgcat gcaagatcta tctagaaggg ccccactagt gacgcgtact cgagggtcga
     2281 cggtatcgat aagcttgata tacactccgc tagcgctgat gtccggcggt gcttttgccg
     2341 ttacgcacca ccccgtcagt agctgaacag gagggacagc tgatagaaac agaagccact
     2401 ggagcacctc aaaaacacca tcatacacta aatcagtaag ttggcagcat cacccgacgc
     2461 actttgcgcc gaataaatac ctgtgacgga agatcacttc gcagaataaa taaatcctgg
     2521 tgtccctgtt gataccggga agccctgggc caacttttgg cgaaaatgag acgttgatcg
     2581 gcacgtaaga ggttccaact ttcaccataa tgaaataaga tcactaccgg gcgtattttt
     2641 tgagttatcg agattttcag gagctaagga agctaaaatg gagaaaaaaa tcactggata
     2701 taccaccgtt gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt
     2761 tgctcaatgt acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt
     2821 aaagaaaaat aagcacaagt tttatccggc ctttattcac attcttgccc gcctgatgaa
     2881 tgctcatccg gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt
     2941 tcacccttgt tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga
     3001 ataccacgac gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg
     3061 tgaaaacctg gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa
     3121 tccctgggtg agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc
     3181 ccccgttttc accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc
     3241 gattcaggtt catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt
     3301 acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt ttttaaggca gttattggtg
     3361 cccttaaacg cctggttgct acgcctgaat aagtgataat aagcggatga atggcagaaa
     3421 ttcgaaagca aattcgaccc ggtcgtcggt tcagggcagg gtcgttaaat agccgcttat
     3481 gtctattgct ggtttaccgg tttattgact accggaagca gtgtgaccgt gtgcttctca
     3541 aatgcctgag gccagtttgc tcaggctctc cccgtggagg taataattga cgatatgatc
     3601 atttattctg cctcccagag cctgataaaa acggttagcg cttcgttaat acagatgtag
     3661 gtgttccaca gggtagccag cagcatcctg cgatgcagat catcgaattc ctgcagccaa
     3721 gctagaccta ggccttaaga tcctttttaa cccatcacat atacctgccg ttcactatta
     3781 tttagtgaaa tgagatatta tgatattttc tgaattgtga ttaaaaaggc aactttatgc
     3841 ccatgcaaca gaaactataa aaaatacaga gaatgaaaag aaacagatag attttttagt
     3901 tctttaggcc cgtagtctgc aaatcctttt atgattttct atcaaacaaa agaggaaaat
     3961 agaccagttg caatccaaac gagagtctaa tagaatgagg tcgaaaagta aatcgcgcgg
     4021 gtttgttact gataaagcag gcaagaccta aaatgtgtaa agggcaaagt gtatactttg
     4081 gcgtcacccc ttacatattt taggtctttt tttattgtgc gtaactaact tgccatcttc
     4141 aaacaggagg gctggaagaa gcagaccgct aacacagtac ataaaaaagg agacatgaac
     4201 gatgaacatc aaaaagtttg caaaacaagc aacagtatta acctttacta ccgcactgct
     4261 ggcaggaggc gcaactcaag cgtttgcgaa agaaacgaac caaaagccat ataaggaaac
     4321 atacggcatt tcccatatta cacgccatga tatgctgcaa atccctgaac agcaaaaaaa
     4381 tgaaaaatat caagttcctg aattcgattc gtccacaatt aaaaatatct cttctgcaaa
     4441 aggcctggac gtttgggaca gctggccatt acaaaacgct gacggcactg tcgcaaacta
     4501 tcacggctac cacatcgtct ttgcattagc cggagatcct aaaaatgcgg atgacacatc
     4561 gatttacatg ttctatcaaa aagtcggcga aacttctatt gacagctgga aaaacgctgg
     4621 ccgcgtcttt aaagacagcg acaaattcga tgcaaatgat tctatcctaa aagaccaaac
     4681 acaagaatgg tcaggttcag ccacatttac atctgacgga aaaatccgtt tattctacac
     4741 tgatttctcc ggtaaacatt acggcaaaca aacactgaca actgcacaag ttaacgtatc
     4801 agcatcagac agctctttga acatcaacgg tgtagaggat tataaatcaa tctttgacgg
     4861 tgacggaaaa acgtatcaaa atgtacagca gttcatcgat gaaggcaact acagctcagg
     4921 cgacaaccat acgctgagag atcctcacta cgtagaagat aaaggccaca aatacttagt
     4981 atttgaagca aacactggaa ctgaagatgg ctaccaaggc gaagaatctt tatttaacaa
     5041 agcatactat ggcaaaagca catcattctt ccgtcaagaa agtcaaaaac ttctgcaaag
     5101 cgataaaaaa cgcacggctg agttagcaaa cggcgctctc ggtatgattg agctaaacga
     5161 tgattacaca ctgaaaaaag tgatgaaacc gctgattgca tctaacacag taacagatga
     5221 aattgaacgc gcgaacgtct ttaaaatgaa cggcaaatgg tacctgttca ctgactcccg
     5281 cggatcaaaa atgacgattg acggcattac gtctaacgat atttacatgc ttggttatgt
     5341 ttctaattct ttaactggcc catacaagcc gctgaacaaa actggccttg tgttaaaaat
     5401 ggatcttgat cctaacgatg taacctttac ttactcacac ttcgctgtac ctcaagcgaa
     5461 aggaaacaat gtcgtgatta caagctatat gacaaacaga ggattctacg cagacaaaca
     5521 atcaacgttt gcgccaagct tcctgctgaa catcaaaggc aagaaaacat ctgttgtcaa
     5581 agacagcatc cttgaacaag gacaattaac agttaacaaa taaaaacgca aaagaaaatg
     5641 ccgatatcct attggcattt tcttttattt cttatcaaca taaaggtgaa tcccatatga
     5701 actatataaa agcaggcaaa tggctaaccg tattcctaac cttttggtaa tgactccaac
     5761 ttattgatag tgttttatgt tcagataatg cccgatgact ttgtcatgca gctccaccga
     5821 ttttgagaac gacagcgact tccgtcccag ccgtgccagg tgctgcctca gattcaggtt
     5881 atgccgctca attcgctgcg tatatcgctt gctgattacg tgcagctttc ccttcaggcg
     5941 ggattcatac agcggccagc catccgtcat ccatatcacc acgtcaaagg gtgacagcag
     6001 gctcataaga cgccccagcg tcgccatagt gcgttcaccg aatacgtgcg caacaaccgt
     6061 cttccggaga ctgtcatacg cgtaaaacag ccagcgctgg cgcgatttag ccccgacata
     6121 gccccactgt tcgtccattt ccgcgcagac gatgacgtca ctgcccggct gtatgcgcga
     6181 ggttaccgac tgcggcctga gttttttaag tgacgtaaaa tcgtgttgag gccaacgccc
     6241 ataatgcggg ctgttgcccg gcatccaacg ccattcatgg ccatatcaat gattttctgg
     6301 tgcgtaccgg gttgagaagc ggtgtaagtg aactgcagca atggcaacaa cgttgcgcaa
     6361 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6421 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     6481 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     6541 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     6601 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6661 ccaagtttac tcatatatac tttagattga tttatggtgc actctcagta caatctgctc
     6721 tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct
     6781 gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     6841 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     6901 tcatcaccga aacgcgcgag gcagcaagga gatggcgccc aacagtcccc cggccacggg
     6961 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     7021 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     7081 gccggccacg atgcgtccgg cgtagag
//
