LOCUS       40924_33472        4482 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNQ705-1, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4482)
  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 4482)
  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 4482)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4482)
  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     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..4482
                     /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="AGO64132.1"
                     /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        2574..2676
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             2677..3333
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
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 aatcccggga gagctcaggt
     2221 tacccgcatg caagatctat ctagaagggc cccactagtg acgcgtactc gagggtcgac
     2281 ggtatcgata agcttgatat acactccgct agcgctgatg tccggcggtg cttttgccgt
     2341 tacgcaccac cccgtcagta gctgaacagg agggacagct gatagaaaca gaagccactg
     2401 gagcacctca aaaacaccat catacactaa atcagtaagt tggcagcatc acccgacgca
     2461 ctttgcgccg aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt
     2521 gtccctgttg ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg
     2581 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
     2641 gagttatcga gattttcagg agctaargaa gctaaaatgg agaaaaaaat cactggatat
     2701 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
     2761 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
     2821 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
     2881 gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     2941 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     3001 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     3061 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     3121 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     3181 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     3241 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     3301 caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc
     3361 ccttaaacgc ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat
     3421 tcgaaagcaa attcgacccg gtcgtcggtt cagggcaggg tcgttaaata gccgcttatg
     3481 tctattgctg gtttaccggt ttattgacta ccggaagcag tgtgaccgtg tgcttctcaa
     3541 atgcctgagg ccagwttgct cagctctccc gtggaggtaa taattgacga tatgatcatt
     3601 tattctgcct cccagagcct gataaaaacg gttagcgctt cgttaataca gatgtaggtg
     3661 ttccacaggg tagccagcag catcctgcga tgcagatcat cgaattcctg cagcaatggc
     3721 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3781 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3841 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3901 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3961 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4021 ttggtaactg tcagaccaag tttactcata tatactttag attgatttat ggtgcactct
     4081 cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt
     4141 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
     4201 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     4261 cagaggtttt caccgtcatc accgaaacgc gcgaggcagc aaggagatgg cgcccaacag
     4321 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     4381 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     4441 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag ag
//
