LOCUS       V005231                 5611 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005231
VERSION     V005231
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5611)
  AUTHORS   Toda H, Koyanagi T, Enomoto T, Itoh N.
  TITLE     Characterization of two cryptic plasmids from Kocuria palustris
            IPUFS-1 and construction of novel Escherichia coli-Kocuria shuttle
            vector for biocatalysis
  JOURNAL   J. Biosci. Bioeng. 124 (3), 255-262 (2017)
   PUBMED   28495560
REFERENCE   2  (bases 1 to 5611)
  AUTHORS   Itoh N, Toda H, Koyanagi T, Enomoto T.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-DEC-2016) Contact:Nobuya Itoh Toyama Prefectural
            University, Biotechnology Research Center; 5180 Kurokawa, Imizu,
            Toyama 939-0398, Japan URL :http://www.pu-toyama.ac.jp/BR/itoh/
REFERENCE   3  (bases 1 to 5611)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5611)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J. Biosci.
            Bioeng."; date: "2017"; volume: "124"; issue: "3"; pages: "255-262"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (12-DEC-2016) Contact:Nobuya Itoh Toyama Prefectural University,
            Biotechnology Research Center"; volume: " 5180 Kurokawa, Imizu,
            Toyama 939-0398, Japan URL :http"; pages:
            "//www.pu-toyama.ac.jp/BR/itoh"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          3999..5605
                     /plasmid="pKPAL1"
                     /strain="IPUFS-1"
                     /mol_type="other DNA"
                     /db_xref="taxon:71999"
                     /organism="Kocuria palustris"
     protein_bind    39..60
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        75..105
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    113..129
                     /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)."
     primer_bind     137..153
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     misc_feature    163..219
                     /label="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(223..239)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      452..907
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        1189..1293
                     /label="AmpR promoter"
     CDS             1294..2151
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      2325..2913
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             3141..3944
                     /gene="aph"
                     /label="Aminoglycoside 3'-phosphotransferase"
                     /note="Aminoglycoside 3'-phosphotransferase from
                     Streptomyces fradiae. Accession#: P00555"
     CDS             4131..5006
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein"
                     /label="repA"
                     /protein_id="BAX51299.1"
                     /translation="MTTAECVQRWDQMWLPLWPLASDDLHAGVYRQARGEAMHRRYIET
                     NPRALSNLLVVDIDHEDAALRTMWNRQAWWPNAVVENPSNGHAHAVWALNEPITRTEYA
                     RRKPLAFAAAVTEGLRRSVDGDTGYSGLLTKNPAHDDWDALWCNSDRLYDLDELAEHLT
                     EGGYMPPVSWKRSKRRNSVGLGRNCSIFETARTWAYREIRNHWFDSAGLHDAISAHVHE
                     LNAEFPEPLPASEARAIAASIHRWITTKSRMWADGPAVYEATFSTIQAARGRKRVERTK
                     ALLEGWQPNG"
     gene            4131..5006
                     /gene="repA"
                     /label="repA"
     CDS             4999..5307
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein"
                     /label="repB"
                     /protein_id="BAX51300.1"
                     /translation="MGDRYDRKGSSISEMSRGTGLSPATIKRWTSRSRDEWLQQKADER
                     EAIRAFHDDEGHSWPQTAKHFRLDVSTVKRRAYRAREERKQEIAEQLQPPLPFPTNS"
     gene            4999..5307
                     /gene="repB"
                     /label="repB"
ORIGIN
        1 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
       61 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
      121 ggataacaat ttcacacagg aaacagctat gaccatgatt acgaattcga gctcggtacc
      181 cggggatcct ctagagtcga cctgcaggca tgcaagcttg gcactggccg tcgttttaca
      241 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
      301 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
      361 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
      421 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
      481 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      541 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      601 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      661 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      721 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
      781 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
      841 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
      901 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
      961 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     1021 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     1081 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     1141 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     1201 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     1261 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     1321 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     1381 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1441 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1501 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1561 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1621 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1681 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     1741 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     1801 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     1861 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     1921 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     1981 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2041 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2101 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     2161 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     2221 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     2281 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2341 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2401 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2461 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2521 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2581 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2641 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2701 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2761 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2821 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2881 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttagatcc
     2941 tgaacttttg ctttgccacg gaacggtctg cgttgtcggg aagatgcgtg atctgatcct
     3001 tcaactcagc aaaagttcga tttattcaac aaagccacgt tgtgtctcaa aatctctgat
     3061 gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct gcttacataa
     3121 cccgttggag gaggctccaa atggacgact ccacgctgcg ccgcaagtac ccccaccacg
     3181 aatggcacgc ggtgaacgag ggcgactccg gcgccttcgt gtaccagctg accggcggcc
     3241 cggagccgca gccggagctg tacgccaaga tcgccccccg cgccccggag aactccgcct
     3301 tcgacctctc cggcgaggcc gatcgcctcg aatggctcca ccgtcatggt atcccggtgc
     3361 cccgcgtggt ggagcgcggc gccgacgaca ccgccgcctg gctcgtgacg gaggccgtgc
     3421 ccggcgtggc cgcggcggag gagtggccgg agcatcagcg cttcgccgtg gtggaggcta
     3481 tggctgaact ggcccgcgcg ctccatgaac tccccgtgga agattgcccc tccgaccgcc
     3541 gcctggacgc cgccgtggcc gaggcgcgcc gcaacgtcgc cgagggcctc gtggatctgg
     3601 acgacctgca ggaggagcgc gccggctgga ccggcgacca gctgctggcc gagctggacc
     3661 gcacgcggcc cgagaaggaa gacctcgtgg tgtgccacgg cgatctgtgc cccaacaacg
     3721 tgctgctgga ccccggcacc tgccgcgtga ccggcgtgat cgacgtgggc cgcctgggcg
     3781 tggccgaccg ccacgccgac atcgccctgg ccgcccgcga gctggagatc gacgaagacc
     3841 cgtggttcgg cccggcctac gccgagcgct ttctggaacg ctacggcgcc caccgcgtgg
     3901 ataaggagaa gctggccttc taccagctcc tggatgagtt cttctgagcg cctctgttcc
     3961 tcggacgctc ggttgctcga cctcgattcg tcagatcttg cggctgaacg aacacgggca
     4021 ccccaggggc aaagcggact gcgctgcgcg cgggtgggtg gatgcccgaa acatctgact
     4081 tggctaagag ccactttgca ccgggtcatg tttgtgggag agtcgcacac atgacgacgg
     4141 cggagtgcgt gcagcggtgg gatcagatgt ggttgccgct gtggccgttg gcctccgacg
     4201 acctgcacgc gggtgtgtac cggcaggcgc gcggtgaggc gatgcatcgt cgctacatcg
     4261 agacgaaccc gcgtgcgctg agcaacctgc tcgtggtgga catcgaccac gaggacgccg
     4321 cgctgcgcac gatgtggaac cgacaggcct ggtggcccaa cgcggtcgtg gagaacccgt
     4381 ccaacggcca cgcgcacgcc gtctgggcgc tcaacgaacc gatcacccgc acggagtacg
     4441 cccgtcgcaa gccgctggcg ttcgcagcag ctgtgaccga ggggctgcgc cgctccgtgg
     4501 acggcgacac gggctactcc gggctgctga ccaagaaccc cgcacacgac gactgggacg
     4561 cgttgtggtg caacagcgac cggctctacg acctcgacga gctcgctgag cacctcaccg
     4621 agggcgggta catgcctccg gtgtcctgga agcgctccaa gcgccgcaac agcgtcgggc
     4681 tgggccgcaa ctgctcgatc ttcgagaccg cacgcacctg ggcctaccgc gagatccgca
     4741 accactggtt cgacagtgcc ggcctccacg acgcgatcag cgcacacgtg cacgaactca
     4801 atgccgagtt ccccgaacca ctgccggcat cagaagcccg agcgatcgcg gccagcatcc
     4861 accgctggat caccaccaag tcccgcatgt gggccgacgg ccccgccgtc tacgaagcca
     4921 cgttctccac catccaagcc gcacgcgggc gcaagcgcgt ggaacgcacc aaggcactct
     4981 tggaggggtg gcagcccaat gggtgatcgg tacgaccgca aggggagttc catctcggag
     5041 atgtcgcgag gaactggtct gagcccagcc acgatcaagc gatggacctc gcgtagtcgg
     5101 gatgagtggc ttcagcagaa agccgacgag cgtgaggcca tccgtgcgtt ccacgacgat
     5161 gaggggcact cctggccgca gacggccaag cacttccggc tcgacgtctc gacggtgaag
     5221 cgccgcgcgt atcgagcgcg tgaggagcgc aaacaggaga tcgccgagca gttgcagcct
     5281 ccgctgccct tccccaccaa ctcctgaacc atcagctatc aaaaacgaaa ttcgcccccg
     5341 ggactccgac ggagccccgg gggcgaattt ccggtatatg gtataatgtc cttagcgatc
     5401 attccaagca gtgccatgcc tgggattccg gtcgcccgag gagatggtcc atatccgtag
     5461 tgctctgtat atacattacg ttatcaggac tgcgacatca gagcaaccgt ggactcccac
     5521 gactgccccg tcgcgacggc ccaagcgggc cgcccctacc gacctcctct cctcggacct
     5581 tcgtcgcgcc gaggagagga ggaacccatg g
//
