LOCUS       40924_33547        5058 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNT562 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5058)
  AUTHORS   Nishida T, Watanabe K, Tachibana M, Shimizu T, Watarai M.
  TITLE     Characterization of the cryptic plasmid pOfk55 from Legionella 
            pneumophila and construction of a pOfk55-derived shuttle vector
  JOURNAL   Plasmid 90, 30-37 (2017)
  PUBMED    28259635
REFERENCE   2  (bases 1 to 5058)
  AUTHORS   Nishida T, Watanabe K, Tachibana M, Shimizu T, Watarai M.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-FEB-2017) Contact:Takashi Nishida Yamaguchi 
            University, The United Graduate School of Veterinary Science; 1677-1
            Yoshida, Yamaguchi, Yamaguchi 753-8515, Japan
REFERENCE   3  (bases 1 to 5058)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5058)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid 90,
            30-37 (2017)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-FEB-2017) Contact:Takashi Nishida Yamaguchi University, The 
            United Graduate School of Veterinary Science; 1677-1 Yoshida, 
            Yamaguchi, Yamaguchi 753-8515, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5058
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      563..572
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1016..1603
                     /codon_start=1
                     /product="putative replication protein"
                     /label=putative replication protein
                     /protein_id="BAX01899.1"
                     /translation="MKKEPIPGKRKNVLAYPENPFWQKTEIKIGSKMVKVSGGKHINIE
                     GESISHSGIHVVKEIDENEFVKIYTKNIKAIFDLKPTAQRVLQYLITELQKTPNADAVY
                     LAWVGAEEYFSENHIKSSRASFHRALSELIQKGFLAESTKPNMFWFNPNLFFNGNRLTF
                     IHEYRKKTIQEIGKEENLVQSDIDKQIQIIFN"
     CDS             complement(1678..2757)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(2758..2833)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        3067..3095
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    3103..3119
                     /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)."
     misc_feature    complement(3166..3273)
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     CDS             3479..4270
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4452..5040
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tcgagggtgg cttacgctgc gcttaaaacc acccatcgcc caaagcgtgg gtcgctgcgt
       61 cagcgtcaag gctaaaccct acgggcgcgc tatgcgcggc cttgaccccg actccgctta
      121 aatgacaatg cacacctttc gcattggaat gctaatttgt aattatttga tttttattaa
      181 tttattttat ttaactcaaa taagttattt gaaatatatt cgactggcta ttcaaaaagc
      241 aaaaaggatt tataaaatac ttatttttgt ctattaacaa accattttta tgcatcacaa
      301 gtgcatcaat gcgctgcata ggtgatgcac ctcctattgt ttttctcagt ttaatcaagg
      361 gtttcacatt aaatcatcga gttcatacga aatgcatcat gggcaacaat cacgattacg
      421 aaaggtgtgc attgtcattt aagcggagtc ggggtcaagg ccgcgcatag cgcgcccgta
      481 gggtttagcc ttgacgctga cgcagcgacc cacgctttgg gcgatgggtg gttttaagcg
      541 cagcgtaagc caccctcgag ccccatggcg gcgaatggct tccccgcaag gggaacggga
      601 agggggattt aaaaagcccc gtaggggcac tcctgagccg ttaggcgatg tcttcttgtt
      661 ttaacatcga aatagtagct aagaaagatg atcgtggctt aaaacgcaaa tttagcgatt
      721 ttttttattt ttaattcaaa tattttctga gcgttagcga agtccttcta ttgtttaaat
      781 tttaccccta ctgaagaatt aagaagagaa atgccacttt gcccagtaaa atcaaccccc
      841 aacgtgtcac gccgtgagac aaacagtctc acagctcatg cgactgtttg tttcaaaaaa
      901 ccatttgaaa atgagaccga atcaaattac catgtttttt tgagacaagt agtatcaagc
      961 aatgacacaa ggaatctcaa aacaaacaaa caaaatgaga caataaggaa aagaaatgaa
     1021 aaaagaacca atcccaggaa aaaggaaaaa tgtattggcg tacccagaaa acccattttg
     1081 gcaaaaaaca gagataaaaa taggatccaa aatggttaaa gtttctggag gaaaacacat
     1141 aaatattgaa ggagaaagta tttctcatag tggaattcac gtcgtaaaag aaattgatga
     1201 aaatgagttt gtaaaaattt acacaaagaa cataaaagca attttcgatt tgaaaccaac
     1261 agctcaaagg gtattacaat atttgataac tgaattacaa aaaacaccaa acgccgacgc
     1321 agtttatttg gcttgggtag gagccgaaga gtacttttca gaaaaccata taaaatcatc
     1381 aagagcatct tttcacagag ctctcagtga attaattcaa aaaggttttt tggccgagag
     1441 caccaagcca aatatgtttt ggtttaatcc aaatctattt tttaacggca atagattaac
     1501 ttttattcat gaatatcgaa agaaaaccat ccaagaaata ggaaaagagg aaaatctagt
     1561 tcaatctgat atcgataaac aaatacaaat aatatttaat taacgccagc aagacgtagc
     1621 ccagcgcgtc ggccagcttg caattcgcgc taacttacat taattgcgtt gcgctcactg
     1681 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     1741 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     1801 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     1861 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     1921 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     1981 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     2041 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     2101 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     2161 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     2221 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     2281 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     2341 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     2401 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     2461 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     2521 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     2581 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     2641 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     2701 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     2761 caccaccctg aattgactct cttgggcgct atcatgccat accgcgaaag gttttgcacc
     2821 attcgatggt gtcaacgtaa atgcatgccg cttcgccttc gcgcggaatt gcaagctgat
     2881 ccggagctta tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc
     2941 tgtggtatgg ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc
     3001 ccgttctgga taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat
     3061 gagctgttga caattaatca tcggctcgta taatgtgtgg aattgtgagc ggataacaat
     3121 ttcacacagg aaacattaat taacgactca ctatagggcg aattgggtac cgggcccccc
     3181 ctcgaggtcg acggtatcga taagcttgat atcgaattcc tgcagcccgg gggatccact
     3241 agttctagag cggccgccac cgcggtggag ctccagcttt tgttattaag ggcttacatg
     3301 acgatagcta gactgggcgg ttttatggac agcaagcgaa ccggaattgc cagctggggc
     3361 gccctctggt aaggttggga agccctgcaa agtaaactgg atggctttct tgccgccaag
     3421 gatctgatgg cgcaggggat caagatctga tcaagagaca ggatgaggat cgtttcgcat
     3481 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     3541 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     3601 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     3661 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     3721 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     3781 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     3841 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     3901 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     3961 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     4021 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     4081 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     4141 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     4201 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     4261 cgagttcttc tgactcgata actgtcagac caagtttact catatatact ttagattgat
     4321 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     4381 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     4441 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     4501 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     4561 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     4621 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     4681 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     4741 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     4801 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     4861 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     4921 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     4981 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     5041 aacgccagca acgcggcc
//
