LOCUS       40924_34619        9375 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PiggyBac donor vector pPIGA3 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9375)
  AUTHORS   Mitsutake H, Kobayashi J.
  TITLE     piggyBac-mediated stable transformation of cultured Bombyx mori 
            cells using in vitro synthesized transposase mRNA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 9375)
  AUTHORS   Mitsutake H, Kobayashi J.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-OCT-2010) Contact:Jun Kobayashi Yamaguchi University, 
            Faculty of Agriculture; Yoshida 1677-1, Yamaguchi, Yamaguchi 
            753-8515, Japan URL 
            :http://www.agr.yamaguchi-u.ac.jp/~member/kobayashi/pe.html
REFERENCE   3  (bases 1 to 9375)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 9375)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-OCT-2010) Contact:Jun Kobayashi Yamaguchi University, Faculty of
            Agriculture"; volume: " Yoshida 1677-1, Yamaguchi, Yamaguchi 
            753-8515, Japan URL :http"; pages: 
            "//www.agr.yamaguchi-u.ac.jp/~member/kobayashi/pe.htm"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        96..200
                     /label=AmpR promoter
     CDS             201..1058
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1232..1820
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2144..2174
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   2375..2410
                     /label=piggyBac 5' inverted terminal repeat
                     /note="piggyBac 5' inverted terminal repeat"
     promoter        3131..3583
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     CDS             3682..4278
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    4335..4640
                     /gene="PGK"
                     /label=PGK polyadenylation sequence
                     /note="PGK polyadenylation sequence"
     gene            4335..4640
                     /gene="PGK"
                     /label=PGK
     gene            4658..5313
                     /gene="A3"
                     /label=A3
     regulatory      4658..4784
                     /gene="A3"
                     /label=actin A3
                     /note="actin A3"
                     /regulatory_class="promoter"
     exon            4785..4853
                     /gene="A3"
                     /note="actin A3"
     intron          4854..5313
                     /gene="A3"
                     /note="actin A3"
     CDS             5343..6059
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    6185..6306
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(6313..6768)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6795..6899
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6942..7239
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     misc_feature    8105..8128
                     /note="piggyBac transposase polyadenylation sequence; 
                     transposase"
     repeat_region   8230..8292
                     /label=piggyBac 3' inverted terminal repeat
                     /note="piggyBac 3' inverted terminal repeat"
     primer_bind     complement(8981..8997)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgaatttcga cgctcgcgcg acttggtttg ccattcttta gcgcgcgtcg
     2281 cgtcacacag cttggccaca atgtggtttt tgtcaaacga agattctatg acgtgtttaa
     2341 agtttaggtc gagtaaagcg caaatctttt ttaaccctag aaagatagtc tgcgtaaaat
     2401 tgacgcatgc attcttgaaa tattgctctc tctttctaaa tagcgcgaat ccgtcgctgt
     2461 gcatttagga catctcagtc gccgcttgga gctcggctga ggcgtgcttg tcaatgcggt
     2521 aagtgtcact gattttgaac tataacgacc gcgtgagtca aaatgacgca tgattatctt
     2581 ttacgtgact tttaagattt aactcatacg ataattaata ttgttatttc atgttctact
     2641 tacgtgataa cttattatat atatattttc ttgttataga tatcgtgact aatatataat
     2701 aaaatgggat gttctttaga cgatgagcat atcctctctg ctcttctgca aggcgatgac
     2761 gagcttgttg gtgaggattc tgacagtgaa atatcagatc acgtaagtga agacgtccag
     2821 agcgatacag aagaagcgtt tatagatgag gtacatgaag tgtcagccaa cgtcaagcgt
     2881 agtgaaatat tagacgaaca aaatgttatt gaacaaccag gttcttcatt ggcttctaac
     2941 agaatcttga ccttgccaca gaggactatt agaggtaaga ataaacattg ttggtcaact
     3001 tcaaagtcca cgagcggtag ccgagtctct gcactgaaca ttgtcagatc tctagagtct
     3061 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgaattaat
     3121 tcccatcccc ctagaatccc aaaacaaact ggttattgtg gtaggtcatt tgtttggcag
     3181 aaagaaaact cgagaaattt ctctggccgt tattcgttat tctctctttt ctttttgggt
     3241 ctctccctct ctgcactaat gctctctcac tctgtcacac agtaaacggc atactgctct
     3301 cgttggttcg agagagcgcg cctcgaatgt tcgcgaaaag agcgccggag tataaataga
     3361 ggcgcttcgt ctacggagcg acaattcaat tcaaacaagc aaagtgaaca cgtcgctaag
     3421 cgaaagctaa gcaaataaac aagcgcagct gaacaagcta aacaatctgc agtaaagtgc
     3481 aagttaaagt gaatcaatta aaagtaacca gcaaccaagt aaatcaactg caactactga
     3541 aatctgccaa gaagtaatta ttgaatacaa gaagagaact ctgggggatc tgatcaagag
     3601 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggac
     3661 ctgcaggcat gcaagcttac catgaccgag tacaagccca cggtgcgcct cgccacccgc
     3721 gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg
     3781 cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc
     3841 ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg
     3901 gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg
     3961 cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg
     4021 gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac
     4081 caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc
     4141 gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg
     4201 ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg
     4261 acccgcaagc ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc
     4321 gcacgacccc atgcatcgat gatatcagat ccccgggatg cagaaattga tgatctatta
     4381 aacaataaag atgtccacta aaatggaagt ttttcctgtc atactttgtt aagaagggtg
     4441 agaacagagt acctacattt tgaatggaag gattggagct acgggggtgg gggtggggtg
     4501 ggattagata aatgcctgct ctttactgaa ggctctttac tattgcttta tgataatgtt
     4561 tcatagttgg atatcataat ttaaacaagc aaaaccaaat taagggccag ctcattcctc
     4621 ccactcatga tctatagatc tcgagctcaa gcttgatcgc gttaccatat atggtgacaa
     4681 aaactgagtc agcccgcgat tggtggaaaa acaaactgga gccgatactg tgtaaattgt
     4741 gataacggct cttttatata gtttatcctc acgagtcggt tctcatttac taaggtgtgc
     4801 tcgaacagtg cgcattcgca tctacgtact tgtcacttat ttaataatac tatgtaagtt
     4861 ttaattttaa aattgcgaaa gaaaaaaaaa catatttatt tatttgtaaa atttgaattt
     4921 cgaaggttct ccgtcccttt acctttaagt attacatatg tttgagtgtt tttttttttt
     4981 aataatacgc taatgataac gtgttacgtt acataattgt tgcataacta gtgaagtgaa
     5041 attttttata aaaaaaaaca tttttcggaa tttagtgtac tgcagatgtt aataaacact
     5101 actaaataag aaataagttt attggacgca catttcaaag tgtccactcg catcgatcaa
     5161 ttcggaaaca gaaattggga acagtgaatt atgaatctta tacagttttc tttaacgtca
     5221 ctaaatagat ggacgcaaat aaatttgtcg tttacttagt ataatgtatg gaatgagaat
     5281 gtagtttgaa ttgttttttt tcttttcttg cagactaatt caaggatcca ccggtcgcca
     5341 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     5401 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     5461 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     5521 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     5581 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     5641 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     5701 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     5761 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     5821 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     5881 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     5941 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     6001 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     6061 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     6121 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     6181 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6241 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6301 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     6361 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     6421 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     6481 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     6541 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     6601 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     6661 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     6721 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     6781 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     6841 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     6901 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     6961 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     7021 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     7081 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     7141 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     7201 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggaaccattg tgggaaccgt
     7261 gcgatcaaac aaacgcgaga taccggaagt actgaaaaac agtcgctcca ggccagtggg
     7321 aacatcgatg ttttgttttg acggacccct tactctcgtc tcatataaac cgaagccagc
     7381 taagatggta tacttattat catcttgtga tgaggatgct tctatcaacg aaagtaccgg
     7441 taaaccgcaa atggttatgt attataatca aactaaaggc ggagtggaca cgctagacca
     7501 aatgtgttct gtgatgacct gcagtaggaa gacgaatagg tggcctatgg cattattgta
     7561 cggaatgata aacattgcct gcataaattc ttttattata tacagccata atgtcagtag
     7621 caagggagaa aaggttcaaa gtcgcaaaaa atttatgaga aacctttaca tgagcctgac
     7681 gtcatcgttt atgcgtaacc gtttagaagc tcctactttg aagagatatt tgcgcgataa
     7741 tatctctaat attttgccaa atgaagtgcc tggtacatca gatgacagta ctgaagagcc
     7801 agtaatgaaa aaacgtactt actgtactta ctgcccctct aaaataaggc gaaaggcaaa
     7861 tgcatcgtgc aaaaaatgca aaaaagttat ttgtcgagag cataatattg atatgtgcca
     7921 aagttgtttc tgactgacta ataagtataa tttgtttcta ttatgtataa gttaagctaa
     7981 ttacttattt tataatacaa catgactgtt tttaaagtac aaaataagtt tatttttgta
     8041 aaagagagaa tgtttaaaag ttttgttact ttatagaaga aattttgagt ttttgttttt
     8101 ttttaataaa taaataaaca taaataaatt gtttgttgaa tttattatta gtatgtaagt
     8161 gtaaatataa taaaacttaa tatctattca aattaataaa taaacctcga tatacagacc
     8221 gataaaacac atgcgtcaat tttacgcatg attatcttta acgtacgtca caatatgatt
     8281 atctttctag ggttaaataa tagtttctaa tttttttatt attcagcctg ctgtcgtgaa
     8341 taccgtatat ctcaacgctg tctgtgagat tgtcgtattc tagccttttt agtttttcgc
     8401 tcatcgactt gatattgtcc gacacatttt cgtcgatttg cgttttgatc aaagacttga
     8461 gcagagacac gttaatcaac tgttcaaatt gatccatatt aacgatatca acccgatgcg
     8521 tatatggtgc gtaaaatata ttttttaacc ctcttatact ttgcactctg cgttaatacg
     8581 cgttcgtgta cagacgtaat catgttttct tttttggata aaactcctac tgagtttgac
     8641 ctcatattag accctcacaa gttgcaaaac gtggcatttt ttaccaatga agaatttaaa
     8701 gttattttaa aaaatttcat cacagattta aagaagaacc aaaaattaaa ttatttaatc
     8761 gaccagttaa tcaacgtgta cacagagcgc aaaaaacacg cagcccgacg tgttggctaa
     8821 aattattaaa tcaacttgtg ttatagtcac gatttgccgt ccaacgtgtt cctcaaaaag
     8881 ttgaagacca acaagtttac ggacactagt taattatttg attttgcccc acttcatttt
     8941 gtgggatcac aattttgtta tattttaaac aaagcttggc actggccgtc gttttacaac
     9001 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     9061 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     9121 gcctgaatgg cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     9181 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc
     9241 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     9301 cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat
     9361 caccgaaacg cgcga
//
