LOCUS       40924_21539        8446 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pGFght-EGFP DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8446)
  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  JOURNAL   Unpublished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 8446)
  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 8446)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8446)
  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            "Unpublished"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..8446
                     /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"
     regulatory      3543..4269
                     /gene="Gf.TEF3"
                     /regulatory_class="promoter"
     gene            3543..4269
                     /gene="Gf.TEF3"
                     /label=Gf.TEF3
     CDS             4270..4986
                     /label=EGFP
                     /note="enhanced GFP"
     regulatory      4990..5786
                     /gene="Gf.TEF3"
                     /regulatory_class="terminator"
     gene            4990..5786
                     /gene="Gf.TEF3"
                     /label=Gf.TEF3
     primer_bind     complement(5804..5820)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5828..5844)
                     /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(5852..5882)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5897..5918)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6206..6794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6968..7825)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7826..7930)
                     /label=AmpR promoter
     primer_bind     8404..8420
                     /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 atattcgaaa tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     4321 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     4381 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     4441 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     4501 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     4561 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     4621 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     4681 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     4741 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     4801 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     4861 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     4921 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     4981 tacaagtgaa cgttgtccgt cgccgttgtt gtttatcctg tctctatcgc tgtgttgttc
     5041 tatccggacg tctctctcgg ctatatatct acgtcttgta ctctgtaccc tagtattacc
     5101 tctacttcac gcatttatga catttcgctt ctttctgttc gaattggagg atgatacgga
     5161 gctagacatt gagcgtgaag cagtgtcctc gatgcaactg cggagcgatt gtcggtgcca
     5221 caggcgcgtt ctgacggctg cgcgctcacc gcaattatta tttgaatgac ggctccttga
     5281 ggcttccaat ctctttgctt ccaacttcca cacttctggt ttcacgacct gcgatgccca
     5341 atgttcatga atacccagtg cattcgttcg agctgttcgc tcgctccgcg acgaacacac
     5401 tcacaccgaa cacgactcag cgaaacacgc ttatagtcgc tgcttgctat atcctggcca
     5461 ttgccattct atggtgcgtt ccctagcctc tgagcttctg cgagtggcgt cgaatttgac
     5521 tcttctcagg catgtgccgt atctctctgc gatcagtgag ttttgcaaag ttcattgtac
     5581 tttccaatgg actaaacgag ttgttttttt ttgcgctcag tatatccttt taagtgagta
     5641 cattcagaaa aacgtaatag gacggtgagg tctcattagc atggtaggct attgactgta
     5701 ggattccatg aggttagtgc atgttcagct ttcgaggtgc ttcctgggct tatgaaccga
     5761 ctgcagatga gccatgcaat cgcaggagct tggcgtaatc atggtcatag ctgtttcctg
     5821 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5881 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5941 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     6001 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     6061 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     6121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     6181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     6241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     6301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     6361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     6421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     6481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     6541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     6601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     7021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     7081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     7141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     7201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     7261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     7321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     7381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     7441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     7501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     7561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     7621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7921 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     7981 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     8041 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     8101 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     8161 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     8221 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     8281 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     8341 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     8401 gttgtaaaac gacggccagt gaattcgagc tcggtacccg gggatc
//
