LOCUS       V005230                 5614 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005230
VERSION     V005230
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 5614)
  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 5614)
  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 5614)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5614)
  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..5614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          4002..5608
                     /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..3947
                     /gene="tsnR"
                     /label="23S rRNA (adenosine(1067)-2'-O)-methyltransferase"
                     /note="23S rRNA (adenosine(1067)-2'-O)-methyltransferase
                     from Streptomyces azureus. Accession#: P18644"
     CDS             4134..5009
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein"
                     /label="repA"
                     /protein_id="BAX51303.1"
                     /translation="MTTAECVQRWDQMWLPLWPLASDDLHAGVYRQARGEAMHRRYIET
                     NPRALSNLLVVDIDHEDAALRTMWNRQAWWPNAVVENPSNGHAHAVWALNEPITRTEYA
                     RRKPLAFAAAVTEGLRRSVDGDTGYSGLLTKNPAHDDWDALWCNSDRLYDLDELAEHLT
                     EGGYMPPVSWKRSKRRNSVGLGRNCSIFETARTWAYREIRNHWFDSAGLHDAISAHVHE
                     LNAEFPEPLPASEARAIAASIHRWITTKSRMWADGPAVYEATFSTIQAARGRKRVERTK
                     ALLEGWQPNG"
     gene            4134..5009
                     /gene="repA"
                     /label="repA"
     CDS             5002..5310
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein"
                     /label="repB"
                     /protein_id="BAX51304.1"
                     /translation="MGDRYDRKGSSISEMSRGTGLSPATIKRWTSRSRDEWLQQKADER
                     EAIRAFHDDEGHSWPQTAKHFRLDVSTVKRRAYRAREERKQEIAEQLQPPLPFPTNS"
     gene            5002..5310
                     /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 atgaccgagc tggacaccat cgcgaacccc tccgatcccg
     3181 ccgtgcagcg gatcatcgac gtgaccaaac cctcccgctc gaacatcaag accaccctca
     3241 tcgaggacgt cgagcccctg atgcactcca tcgccgcggg cgtggagttc attgaggtgt
     3301 acgggtccga ctcgtccccg ttcccctcgg aactgctcga tctgtgcggc cgccagaaca
     3361 tccccgtgcg gctgatcgac tcctccatcg tgaaccagct cttcaagggc gagcgcaaag
     3421 ccaagacctt cgggatcgcc cgcgtgcccc gccccgcgcg cttcggcgac attgcctccc
     3481 gccgcggcga cgtggtggtg ctggacggcg tgaaaatcgt gggcaacatc ggcgcgatcg
     3541 tgcgcacgtc cctcgccctc ggcgcctccg gcatcatcct ggtggactcc gacattacgt
     3601 ccatcgcgga ccgccgcctg cagcgcgcct cccgcgggta tgtgttctcc ctgcccgtgg
     3661 tgctgtccgg gcgcgaggag gcgatcgcct tcatccgcga ctccggcatg cagctgatga
     3721 ccctgaaggc cgacggcgac atctcggtga aggagctggg cgacaacccc gaccgcctgg
     3781 cgctgctgtt cggctccgag aagggcggcc cctccgacct gtttgaggag gcctcctccg
     3841 cgtccgtgtc catccccatg atgtcccaga ccgagtccct gaacgtgtcc gtgtccctgg
     3901 gcatcgcgct gcacgagcgc atcgaccgca acctcgccgc gaaccgctga gcgcctctgt
     3961 tcctcggacg ctcggttgct cgacctcgat tcgtcagatc ttgcggctga acgaacacgg
     4021 gcaccccagg ggcaaagcgg actgcgctgc gcgcgggtgg gtggatgccc gaaacatctg
     4081 acttggctaa gagccacttt gcaccgggtc atgtttgtgg gagagtcgca cacatgacga
     4141 cggcggagtg cgtgcagcgg tgggatcaga tgtggttgcc gctgtggccg ttggcctccg
     4201 acgacctgca cgcgggtgtg taccggcagg cgcgcggtga ggcgatgcat cgtcgctaca
     4261 tcgagacgaa cccgcgtgcg ctgagcaacc tgctcgtggt ggacatcgac cacgaggacg
     4321 ccgcgctgcg cacgatgtgg aaccgacagg cctggtggcc caacgcggtc gtggagaacc
     4381 cgtccaacgg ccacgcgcac gccgtctggg cgctcaacga accgatcacc cgcacggagt
     4441 acgcccgtcg caagccgctg gcgttcgcag cagctgtgac cgaggggctg cgccgctccg
     4501 tggacggcga cacgggctac tccgggctgc tgaccaagaa ccccgcacac gacgactggg
     4561 acgcgttgtg gtgcaacagc gaccggctct acgacctcga cgagctcgct gagcacctca
     4621 ccgagggcgg gtacatgcct ccggtgtcct ggaagcgctc caagcgccgc aacagcgtcg
     4681 ggctgggccg caactgctcg atcttcgaga ccgcacgcac ctgggcctac cgcgagatcc
     4741 gcaaccactg gttcgacagt gccggcctcc acgacgcgat cagcgcacac gtgcacgaac
     4801 tcaatgccga gttccccgaa ccactgccgg catcagaagc ccgagcgatc gcggccagca
     4861 tccaccgctg gatcaccacc aagtcccgca tgtgggccga cggccccgcc gtctacgaag
     4921 ccacgttctc caccatccaa gccgcacgcg ggcgcaagcg cgtggaacgc accaaggcac
     4981 tcttggaggg gtggcagccc aatgggtgat cggtacgacc gcaaggggag ttccatctcg
     5041 gagatgtcgc gaggaactgg tctgagccca gccacgatca agcgatggac ctcgcgtagt
     5101 cgggatgagt ggcttcagca gaaagccgac gagcgtgagg ccatccgtgc gttccacgac
     5161 gatgaggggc actcctggcc gcagacggcc aagcacttcc ggctcgacgt ctcgacggtg
     5221 aagcgccgcg cgtatcgagc gcgtgaggag cgcaaacagg agatcgccga gcagttgcag
     5281 cctccgctgc ccttccccac caactcctga accatcagct atcaaaaacg aaattcgccc
     5341 ccgggactcc gacggagccc cgggggcgaa tttccggtat atggtataat gtccttagcg
     5401 atcattccaa gcagtgccat gcctgggatt ccggtcgccc gaggagatgg tccatatccg
     5461 tagtgctctg tatatacatt acgttatcag gactgcgaca tcagagcaac cgtggactcc
     5521 cacgactgcc ccgtcgcgac ggcccaagcg ggccgcccct accgacctcc tctcctcgga
     5581 ccttcgtcgc gccgaggaga ggaggaaccc atgg
//
