LOCUS       40924_21544        8614 bp DNA     circular SYN 18-DEC-2018
DEFINITION  RNAi vector pGFght-HydA1IR DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8614)
  AUTHORS   Sato M, Kurahashi A, Nishibori K, Fujimori F.
  TITLE     Development of a transformation system for the edible mushroom 
            Grifola frondosa: demonstrating heterologous gene expression and 
            RNAi-mediated gene silencing
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8614)
  AUTHORS   Sato M, Kurahashi A, Nishibori K, Fujimori F.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-FEB-2014) Contact:Masayuki Sato Yukiguni Maitake Co. 
            Ltd., Research and Development; 89 Yokawa, Minamiuonuma, Niigata 
            949-6695, Japan URL :http://www.maitake.co.jp
REFERENCE   3  (bases 1 to 8614)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8614)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-FEB-2014) Contact:Masayuki Sato Yukiguni Maitake Co. Ltd., 
            Research and Development; 89 Yokawa, Minamiuonuma, Niigata 949-6695,
            Japan URL :http://www.maitake.co.jp"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8614
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..1500
                     /gene="Gf.GAPDH"
                     /regulatory_class="promoter"
     gene            1..1500
                     /gene="Gf.GAPDH"
                     /label=Gf.GAPDH
     CDS             1501..2523
                     /label=HygR
                     /note="aminoglycoside phosphotransferase from E. coli"
     regulatory      2527..3526
                     /gene="Gf.GAPDH"
                     /regulatory_class="terminator"
     gene            2527..3526
                     /gene="Gf.GAPDH"
                     /label=Gf.GAPDH
     misc_feature    3527..3542
                     /label=multiple cloning site (BamHI-XbaI-SalI) from pUC19
                     /note="multiple cloning site (BamHI-XbaI-SalI) from pUC19"
     gene            3543..5944
                     /gene="Gf.TEF3"
                     /label=Gf.TEF3
     regulatory      3543..4269
                     /gene="Gf.TEF3"
                     /regulatory_class="promoter"
     misc_feature    4270..4591
                     /gene="Gf.TEF3"
                     /label=Gf.HydA1 sense strand sequence
                     /note="Gf.HydA1 sense strand sequence"
     misc_feature    4592..4821
                     /gene="Gf.TEF3"
                     /note="intron of Gf.GAPDH to serve as the spacer for
                     hairpin formation"
     misc_feature    4822..4829
                     /gene="Gf.TEF3"
                     /label=synthetic sequence for NotI site
                     /note="synthetic sequence for NotI site"
     misc_feature    complement(4830..5151)
                     /label=Gf.HydA1 anti-sense strand sequence
                     /note="Gf.HydA1 anti-sense strand sequence"
     misc_feature    5152..5157
                     /gene="Gf.TEF3"
                     /label=synthetic sequence for SpeI site
                     /note="synthetic sequence for SpeI site"
     regulatory      5158..5944
                     /gene="Gf.TEF3"
                     /regulatory_class="terminator"
     primer_bind     complement(5972..5988)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5996..6012)
                     /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(6020..6050)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6065..6086)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6374..6962)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7136..7993)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7994..8098)
                     /label=AmpR promoter
     primer_bind     8572..8588
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tctgcgcgct tctctcacag ttcacgaagc cgcagataga atgtgtgact ttcatctaca
       61 acgttacact agccgaacag tattccgaac agtcttccga gcagactcag tcctccgagt
      121 ggcaatgtat tctgggcatc ctcagatccc atgcattctc ttcactcaag cggattacct
      181 ggaaaaccgg ccataatgag aagttcgcga atacagtcag gagtgagctg gcctacttgg
      241 atgcgcacgg gatattgcat gtaatccacg gatattgaca gctcgtaatg ggcgttggtg
      301 caggacttcc cgcaggtccg aatgtgtaca tagtgagtga ttcggatgct cctgtggcac
      361 ttctcgggct cacagccgtc ctcagttttg cgaacgccgc ttgttttatt cgccttctca
      421 ttgccttttc tgctgtttgc gttcattacc acacgacgat tttttctgca agctatacaa
      481 ctcatctttc ttcgattaca tgcaagaatc ctgaggtgct tttgcagaaa tctgaccttc
      541 gttcttcgta catacttgtt agccctcgaa ccatcatttt cattctgcat gcacaaaatg
      601 ttgtatttgt aaagtggagg aggctagatt tctctcgaca tatgtatgta gctaagtaaa
      661 actactaccg aagaaactcc cagtcttcta gatcgcctac atcgcgccga agggcgaaat
      721 gtacgactag agctcaaata catcctgtac gtaacctccg ggaagtggca tcttctgaat
      781 tctgaggccg attcggctga aggcgtattg gctcgagaca gggctatgcg tcgggacgtc
      841 gggagcgttc gaagactcca gactgctgat caaggcgctg cgttcgcatt acacacattg
      901 ttcgaacatg tacagagagt ttgacagaca aaacagtaac acttgtttcg agagagacgc
      961 tcgtagctaa acccgatgtc cgaagaggac cctccccacg cgagtccgca agatgaatgc
     1021 gacagttggt tgccacgagc acagagagcg gcaaataccc tcgagtcatc gtacggagct
     1081 tgtcaatcca gcaaatgtac acgagcatgg caggatccaa tcgtaattag gtggctgaga
     1141 tctgaccacc gagaatgtgc gccctagggg atgagtaaac gcacgtttgc gcgtgaatca
     1201 gcgatgatgc tgtacggtgg tgcttagaga tacgaaaagt tgcaagtgaa cgtaaatgga
     1261 ggaaagggac tggttgggaa tattcatgac aagctggcta gaacaagtcg gaaatctagt
     1321 ctgaggcaaa gccaccagcg gagagccgtt cgcggccttg cggtgacagt cgggcaacgg
     1381 ccggaagctg cccggtgtaa tcatccatct tagataacga tcaccacccc accctataag
     1441 acccctctcc atctctgctc ttctccccat ccttcgtctc caaaaccatt atcctcagca
     1501 atgaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac
     1561 agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat
     1621 gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat
     1681 cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt
     1741 ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg
     1801 caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat
     1861 gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga
     1921 atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat
     1981 cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag
     2041 ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc
     2101 tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg
     2161 atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct
     2221 tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg
     2281 cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac
     2341 ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga
     2401 gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc
     2461 tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag
     2521 gaatagggtg tggagtggat aagagaaaga agtttggtgc tgctgtatga ggactttgcg
     2581 ctacttgtag cttgctgata tgcagtcttg cgtgtttgta caaatgtcaa tagaagcatt
     2641 cggatacctc ttggtggtgt ggttgcgttc gtggtgtttg ccgtcgttgg tgggtgttga
     2701 gagcatccat gtcgcgattg gtcgcgatca tcttgtggca atcttactga atggtgtgag
     2761 catgagcagt tgtcggtgtc gctactatcc tcgcacactt ccatctcgtg gcgttcatct
     2821 tgtggcaaac agtccacgct ggagacaagc gggccagtct cctctacacc tgcgacgccc
     2881 tcagtccaac ttgcttagtt ctgacttcgt tctaccgcac aatggctaca tccgtatgcg
     2941 agctattctc gccatttcca tgtttttgtg caatttctcg aattcgcatg cataattagt
     3001 ccacttgcct ctgtcgaatg catcgcctcc gtccatccca atgacgagac agtgctttca
     3061 caatggtaag ctcattcctt tgcaatgggg cttcgtggcc cttgtactca gaccagtcgt
     3121 cactgaggct aatcaccgtg gcataggaat ctctccacat ttcgctgttt tatagacaga
     3181 tgctgtaaca aatagatttg aggaacctcc gaatgacgat ataattcttc acgccaaggt
     3241 ctcttttttt ttttctccct cgctgtcttt gagcctgatg agaatctcgg cgtttacagt
     3301 atggctcaac ctaccagggc catgttcaca cccttcctgc gaacaacctt tccagtttct
     3361 acgctcattt ccggcagggt tataggagac tgttttgcaa atttcgtgat ttctagtagg
     3421 cgatgtcttc atcttcttgg atcaatgact gaacctgtgt ccgaaccaac aaaggaagtt
     3481 cttcggtcgt gtctcgaaga aacgacaaga ctggttcatc taattggatc ctctagagtc
     3541 gacactgggt ggaagctaat agcaagcatt atgaagctgg tggttatgtg cgtgcccttt
     3601 catttacgtc gatcgatggc acgtcgcaaa catgctcacc ggatcccttc tgcagcaatt
     3661 cacaccgctt agtttcgctc agctgaatga gaagcagtga gatggtaagg ccatagctcg
     3721 gacatgaatc ggatgtcatg tagttgcgat gcatgtatac tcgtagttcc gatgtcggat
     3781 acggcgttag caagaagtgc tgtcatttga acgctgcatg atggcccgtg gggttagctt
     3841 ccccgcattc acatcggatg actgagagac ctggaggcgc ccgaggctga gctggatgtg
     3901 accagcgtga caggacacgt gaatacagtg cattttttcg aagtccgcct gcttacacca
     3961 gcgttatata aaccgagact caacaatccg taaccttatc cccactcttt cctttcctct
     4021 tttcttctat tgtcctatag aaggtggctt ttcttcgtct ctgtcatccc tttcaacagc
     4081 ccgctaactt tttgcgcagg ctccggccaa cctagaaata gagtgctcta tcatccttga
     4141 cgaccttttc gcgtcttatc gcattttaca agtaccccga tcatatcatc gttaaaggtg
     4201 attaccacga cacctaatcc tctcactttt gcaggtctga caatgagatc ggtcgcccag
     4261 atattcgaag ttctccaagc tcgccatctt cgctactgcg gccttcgccg ttcttgcggc
     4321 tgccacccct gtccgccgcc aacagtgcac cactggccag ctccagtgct gcgagtctac
     4381 ctccactgcg aacgacccgg ccaccagcga gctcctcggt ctgatcggcg tcgtcatctc
     4441 tgatgtcgac gcactcgtcg gtctcacctg ctcgccgatc tccgtcatcg gcgttggcag
     4501 tggctctgcg tgcaccgcga acccagtgtg ctgtgactcg tcgcccattg gtggactcgt
     4561 ctccatcgga tgtgttccgg ttaacgtctg aacgtttctt tgtaatctta acataggccg
     4621 catatagtct gtttgttatg cgtgcgagta gtctgctacc cgacccgcgc tgttctgaag
     4681 gataactcgt ctttggcggg tgtagcggcg ataatggcgg atgcgttatc atcgacaaaa
     4741 aggactcttc gcgcttggac atcctcctac ccttgttaag ctgatatctt cgatcggtgt
     4801 ctgaccgcgt tccgtttagg tgcggccgct cagacgttaa ccggaacaca tccgatggag
     4861 acgagtccac caatgggcga cgagtcacag cacactgggt tcgcggtgca cgcagagcca
     4921 ctgccaacgc cgatgacgga gatcggcgag caggtgagac cgacgagtgc gtcgacatca
     4981 gagatgacga cgccgatcag accgaggagc tcgctggtgg ccgggtcgtt cgcagtggag
     5041 gtagactcgc agcactggag ctggccagtg gtgcactgtt ggcggcggac aggggtggca
     5101 gccgcaagaa cggcgaaggc cgcagtagcg aagatggcga gcttggagaa cactagtacg
     5161 ttgtccgtcg ccgttgttgt ttatcctgtc tctatcgctg tgttgttcta tccggacgtc
     5221 tctctcggct atatatctac gtcttgtact ctgtacccta gtattacctc tacttcacgc
     5281 atttatgaca tttcgcttct ttctgttcga attggaggat gatacggagc tagacattga
     5341 gcgtgaagca gtgtcctcga tgcaactgcg gagcgattgt cggtgccaca ggcgcgttct
     5401 gacggctgcg cgctcaccgc aattattatt tgaatgacgg ctccttgagg cttccaatct
     5461 ctttgcttcc aacttccaca cttctggttt cacgacctgc gatgcccaat gttcatgaat
     5521 acccagtgca ttcgttcgag ctgttcgctc gctccgcgac gaacacactc acaccgaaca
     5581 cgactcagcg aaacacgctt atagtcgctg cttgctatat cctggccatt gccattctat
     5641 ggtgcgttcc ctagcctctg agcttctgcg agtggcgtcg aatttgactc ttctcaggca
     5701 tgtgccgtat ctctctgcga tcagtgagtt ttgcaaagtt cattgtactt tccaatggac
     5761 taaacgagtt gttttttttt gcgctcagta tatcctttta agtgagtaca ttcagaaaaa
     5821 cgtaatagga cggtgaggtc tcattagcat ggtaggctat tgactgtagg attccatgag
     5881 gttagtgcat gttcagcttt cgaggtgctt cctgggctta tgaaccgact gcagatgagc
     5941 catgcaatcg caggagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     6001 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     6061 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6121 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6181 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     6241 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     6301 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6361 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6421 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6481 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6541 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6601 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6661 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6721 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6781 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6841 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6901 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6961 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     7021 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     7081 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     7141 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     7201 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     7261 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     7321 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     7381 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     7441 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     7501 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     7561 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     7621 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     7681 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     7741 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     7801 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7861 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7921 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7981 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     8041 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     8101 catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     8161 ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa
     8221 acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     8281 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     8341 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     8401 gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt
     8461 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     8521 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     8581 cggccagtga attcgagctc ggtacccggg gatc
//
