LOCUS       40924_5194        6655 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pB-TUB_DEST_GFP DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6655)
  AUTHORS   Irie T, Sakamoto T.
  TITLE     Construction of selectable maker for Pleurotus ostreatus 
            transformation
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6655)
  AUTHORS   Irie T, Sakamoto T.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-MAR-2016) Contact:Toshikazu Irie University of Shiga 
            Prefecture, School of Environmental Science; 2500, Hassaka-cho, 
            Hikone, Shiga Prefecture 522-8533, Japan URL :http://www.usp.ac.jp/
REFERENCE   3  (bases 1 to 6655)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6655)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (19-MAR-2016) Contact:Toshikazu Irie University of Shiga Prefecture,
            School of Environmental Science; 2500, Hassaka-cho, Hikone, Shiga 
            Prefecture 522-8533, Japan URL :http://www.usp.ac.jp/"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6655
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      16..1699
                     /gene="b-tub"
                     /label=beta-tubulin gene in Pleurotus ostreatus
                     /note="beta-tubulin gene in Pleurotus ostreatus"
                     /regulatory_class="promoter"
     gene            16..1699
                     /gene="b-tub"
                     /label=b-tub
     protein_bind    1706..1730
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1751..2767
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     protein_bind    complement(2787..2811)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2816..3532
                     /label=EmGFP
                     /note="Emerald GFP"
     regulatory      3542..3730
                     /gene="sdi1"
                     /label=Ip subunit gene in Pleurotus ostreatus
                     /note="Ip subunit gene in Pleurotus ostreatus"
                     /regulatory_class="terminator"
     gene            3542..3730
                     /gene="sdi1"
                     /label=sdi1
     promoter        complement(3804..3822)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3840..3856)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3864..3880)
                     /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        complement(3888..3918)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3933..3954)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4242..4830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5004..5861)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5862..5966)
                     /label=AmpR promoter
     rep_origin      complement(6044..6499)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gggcgaattg ggccccgtga tgctgttggg aggaaactaa atttgcgagc taccagaagc
       61 tgcaagtcgt gcgactcgtg cttccaaagg gtacctgtga ataaggcatc gcttgatcga
      121 accatcctgc acgttgatgg agcccgcagg gcctaggaag gcctagggat tatcggttgg
      181 tgtgaaacca agcttgattt tggaagatgc agaatgcgga ataacgcagg caccagcgaa
      241 acttaatgca tcgaagactg gaagctatta ttacgactag cacggttcaa gtagtgcatg
      301 tccactaccc gtactgctgg gttgcagtaa atctttaatt atggcctgcg cctgtcccac
      361 agcatgtcca acgaatgtcc aacagatgtc aagaacatcc tgtatgcatg tcagtatcac
      421 cgggcgaaga agtgtcggat gctatagaac tctgagcgct gagtcggaag tgccggtgag
      481 cgttgtgtgg ggctggaact ccatccattc aatggccggt ccgcatgtcg tacgtctatg
      541 ttcgccgtca ctgcccaggc ttacacacaa cgctgataag atggccccgg cggtatttac
      601 ttacagcaca tgcgacgtgc gaagtgggac ggtgaactcc atccactcga tctgtgcgga
      661 ggtgatagac tgattgggtg cctaccctga ttgaactggt gataattagt agatcctaga
      721 cgacgagtag agacaggcaa aacccagata cgctttataa tcctctaagc cgctcaaaca
      781 ctctttcgtc gtcatcaccc tgtaccgcga gagtcctcga tgttgcccag ggtctgagat
      841 gtagtgggag ttgtttgaga aagtgcggga tctatggaca ttggacatgg gtacgattgc
      901 catttgcccg acactcagat tcagatcgat gccttgcatg actgtcacat tagccgcaag
      961 acccgactac cagttcagcc tggggtgaga ctctctagga tctaggtaag tatcatactt
     1021 tcagagtagc gccacgataa gtcaacagcc acttacagtt catcgctact cactagttac
     1081 ttaagcgccc tatacagcgc ttattttttg tttgaaagtt gagggcatag tctcgagagc
     1141 gatgtcagac aagttacaag ctaaacagta agcgcgtgct gttgtcaaca tgcctcttct
     1201 gctgcggtgt gcatgacatt ctgaatcagt ggcaatcact cgattgagtg aactccaata
     1261 gcacacacgt gaaaaagact gacggagatg atccacaaga gtaataacga cgtgtattac
     1321 cctgtcgaat tgcgcgcgac ctcgttgaat gttcgtgtac gatggtcatg atgtcgactc
     1381 ttgttatgta cagtcttagt atgatcctgc gctatgacca gctgcggtca accccgattg
     1441 ttcaagcgga gctctaaaat ttgcttgccc tcgacctgaa cacgttccga ttaattgtct
     1501 cctccttctt cacgaccaac aaagccaatc catcctccat cctcatctat gtaagttgtt
     1561 ctaactccac tttcatattt gctctggcga ttcgtgcaag aaagacgcgt tgtgcgatgg
     1621 aggaaaaacg cgatcttgcg cgcattccaa aacgtaccat tcctgagata gaacgcgact
     1681 aacttctttt ccatgcagaa ttatcacaag tttgtacaaa aaagcaggct ccgcggccgc
     1741 ccccttcacc atgcctgaac tcaccgcgac gtctgtcgag aagtttctga tcgaaaagtt
     1801 cgacagcgtc tccgacctga tgcagctctc ggagggcgaa gaatctcgtg ctttcagctt
     1861 cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc tgcgccgatg gtttctacaa
     1921 agatcgttat gtttatcggc actttgcatc ggccgcgctc ccgattccgg aagtgcttga
     1981 cattggggaa ttcagcgaga gcctgaccta ttgcatctcc cgccgtgcac agggtgtcac
     2041 gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg cagccggtcg cggaggccat
     2101 ggatgcgatc gctgcggccg atcttagcca gacgagcggg ttcggcccat tcggaccgca
     2161 aggaatcggt caatacacta catggcgtga tttcatatgc gcgattgctg atccccatgt
     2221 gtatcactgg caaactgtga tggacgacac cgtcagtgcg tccgtcgcgc aggctctcga
     2281 tgagctgatg ctttgggccg aggactgccc cgaagtccgg cacctcgtgc acgcggattt
     2341 cggctccaac aatgtcctga cggacaatgg ccgcataaca gcggtcattg actggagcga
     2401 ggcgatgttc ggggattccc aatacgaggt cgccaacatc ttcttctgga ggccgtggtt
     2461 ggcttgtatg gagcagcaga cgcgctactt cgagcggagg catccggagc ttgcaggatc
     2521 gccgcggctc cgggcgtata tgctccgcat tggtcttgac caactctatc agagcttggt
     2581 tgacggcaat ttcgatgatg cagcttgggc gcagggtcga tgcgacgcaa tcgtccgatc
     2641 cggagccggg actgtcgggc gtacacaaat cgcccgcaga agcgcggccg tctggaccga
     2701 tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc cccagcactc gtccgagggc
     2761 aaaggaatag aagggtgggc gcgccgaccc agctttcttg tacaaagtgg tgatcatggt
     2821 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2881 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2941 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     3001 gaccaccttg acctacggcg tgcagtgctt cgcccgctac cccgaccaca tgaagcagca
     3061 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3121 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3181 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3241 ggagtacaac tacaacagcc acaaggtcta tatcaccgcc gacaagcaga agaacggcat
     3301 caaggtgaac ttcaagaccc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3361 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3421 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3481 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaggtac
     3541 ctcacaagtt aacagccacg gattaaaagc atcgagtcag aggcagattt ctttcctgta
     3601 gcagttgaca gttctttcca cttcatcata cagtgtccat cacgagcatc aaatcatatt
     3661 cattctataa catccacttg ttcttgagcc tgtctgaggt aggtataccc tcggtttcac
     3721 ttgcgatgct atcactagtg cggccgcctg caggtcgacc atatgggaga gctcccaacg
     3781 cgttggatgc atagcttgag tattctatag tgtcacctaa atagcttggc gtaatcatgg
     3841 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3901 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3961 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     4021 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     4081 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4141 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     4201 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4261 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4321 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4381 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4441 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4501 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4561 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4621 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4681 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4741 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4801 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4861 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4921 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4981 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     5041 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     5101 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     5161 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     5221 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     5281 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5341 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5401 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5461 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5521 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5581 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5641 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5701 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5761 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5821 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5881 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5941 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg
     6001 tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat
     6061 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     6121 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     6181 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     6241 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     6301 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     6361 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     6421 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     6481 gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     6541 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     6601 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccag
//
