LOCUS       40924_34624        8127 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PiggyBac donor vector pPIGA3Fluc DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8127)
  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  JOURNAL   Unpublished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 8127)
  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 8127)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 8127)
  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            "Unpublished"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..8127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   556..591
                     /label=piggyBac 5' inverted terminal repeat
                     /note="piggyBac 5' inverted terminal repeat"
     promoter        1313..1765
                     /label=hsp70 promoter
                     /note="Drosophila melanogaster hsp70Bb promoter"
     CDS             1864..2460
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    2517..2822
                     /gene="PGK"
                     /label=PGK polyadenylation sequence
                     /note="PGK polyadenylation sequence"
     gene            2517..2822
                     /gene="PGK"
                     /label=PGK
     gene            2840..3495
                     /gene="A3"
                     /label=A3
     regulatory      2840..2966
                     /gene="A3"
                     /label=actin A3
                     /note="actin A3"
                     /regulatory_class="promoter"
     exon            2967..3035
                     /gene="A3"
                     /note="actin A3"
     intron          3036..3495
                     /gene="A3"
                     /note="actin A3"
     CDS             3525..5174
                     /label=luciferase
                     /note="firefly luciferase"
     misc_feature    5395..5418
                     /note="piggyBac transposase polyadenylation sequence; 
                     transposase"
     repeat_region   5520..5582
                     /label=piggyBac 3' inverted terminal repeat
                     /note="piggyBac 3' inverted terminal repeat"
     primer_bind     complement(5906..5922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5930..5946)
                     /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(5954..5984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5999..6020)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6308..6896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7070..7927)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7928..8032)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 acgctcgcgc gacttggttt gccattcttt agcgcgcgtc gcgtcacaca gcttggccac
      481 aatgtggttt ttgtcaaacg aagattctat gacgtgttta aagtttaggt cgagtaaagc
      541 gcaaatcttt tttaacccta gaaagatagt ctgcgtaaaa ttgacgcatg cattcttgaa
      601 atattgctct ctctttctaa atagcgcgaa tccgtcgctg tgcatttagg acatctcagt
      661 cgccgcttgg agctcccgtg aggcgtgctt gtcaatgcgg taagtgtcac tgattttgaa
      721 ctataacgac cgcgtgagtc aaaatgacgc atgattatct tttacgtgac ttttaagatt
      781 taactcatac gataattaat attgttattt catgttctac ttacgtgata acttattata
      841 tatatatttt cttgttatag atatcgtgac taatatataa taaaatgggt agttctttag
      901 acgatgagca tatcctctct gctcttctgc aaagcgatga cgagcttgtt ggtgaggatt
      961 ctgacagtga aatatcagat cacgtaagtg aagacgtcca gagcgataca gaagaagcgt
     1021 ttatagatga ggtacatgaa gtgtcagcca acgtcaagcg tagtgaaata ttagacgaac
     1081 aaaatgttat tgaacaacca ggttcttcat tggcttctaa cagaatcttg accttgccac
     1141 agaggactat tagaggtaag aataaacatt gttggtcaac ttcaaagtcc acgagcggta
     1201 gccgagtctc tgcactgaac attgtcagat cctctagagt ctatgcggtg tgaaataccg
     1261 cacagatgcg taaggagaaa ataccgcatc aggcgaatta attcccatcc ccctagaatc
     1321 ccaaaacaaa ctggttattg tggtaggtca tttgtttggc agaaagaaaa ctcgagaaat
     1381 ttctctggcc gttattcgtt attctctctt ttctttttgg gtctctccct ctctgcacta
     1441 atgctctctc actctgtcac acagtaaacg gcatactgct ctcgttggtt cgagagagcg
     1501 cgcctcgaat gttcgcgaaa agagcgccgg agtataaata gaggcgcttc gtctacggag
     1561 cgacaattca attcaaacaa gcaaagtgaa cacgtcgcta agcgaaagct aagcaaataa
     1621 acaagcgcag ctgaacaagc taaacaatct gcagtaaagt gcaagttaaa gtgaatcaat
     1681 taaaagtaac cagcaaccaa gtaaatcaac tgcaactact gaaatctgcc aagaagtaat
     1741 tattgaatac aagaagagaa ctctggggga tctgatcaag agacaggatg aggatcgttt
     1801 cgcatgattg aacaagatgg attgcacgca ggttctccgg acctgcaggc atgcaagctt
     1861 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     1921 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     1981 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     2041 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     2101 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     2161 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     2221 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     2281 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     2341 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     2401 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     2461 tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg
     2521 atgatatcag atccccggga tgcagaaatt gatgatctat taaacaataa agatgtccac
     2581 taaaatggaa gtttttcctg tcatactttg ttaagaaggg tgagaacaga gtacctacat
     2641 tttgaatgga aggattggag ctacgggggt gggggtgggg tgggattaga taaatgcctg
     2701 ctctttactg aaggctcttt actattgctt tatgataatg tttcatagtt ggatatcata
     2761 atttaaacaa gcaaaaccaa attaagggcc agctcattcc tcccactcat gatctataga
     2821 tctcgagctc aagcttgatc gcgttaccat atatggtgac aaaaactgag tcagcccgcg
     2881 attggtggaa aaacaaactg gagccgatac tgtgtaaatt gtgataacgg ctcttttata
     2941 tagtttatcc tcacgagtcg gttctcattt actaaggtgt gctcgaacag tgcgcattcg
     3001 catctacgta cttgtcactt atttaataat actatgtaag ttttaatttt aaaattgcga
     3061 aagaaaaaaa aacatattta tttatttgta aaatttgaat ttcgaaggtt ctccgtccct
     3121 ttacctttaa gtattacata tgtttgagtg tttttttttt ttaataatac gctaatgata
     3181 acgtgttacg ttacataatt gttgcataac tagtgaagtg aaatttttta taaaaaaaaa
     3241 catttttcgg aatttagtgt actgcagatg ttaataaaca ctactaaata aaaaataagt
     3301 ttattggacg cacatttcaa agtgtccact cgcatcgatc aattcggaaa cagaaattgg
     3361 gaacagtgaa ttatgaatct tatacagttt tctttaacgt cactaaatag atggacgcaa
     3421 ataaatttgt cgtttactta gtataatgta tggaatgaga atgtagtttg aattgttttt
     3481 tttcttttct tgcagactaa ttcaaggatc caccggtcgc caccatggaa gacgccaaaa
     3541 acataaagaa aggcccggcg ccattctatc cgctggaaga tggaaccgct ggagagcaac
     3601 tgcataaggc tatgaagaga tacgccctgg ttcctggaac aattgctttt acagatgcac
     3661 atatcgaggt ggacatcact tacgctgagt acttcgaaat gtccgttcgg ttggcagaag
     3721 ctatgaaacg atatgggctg aatacaaatc acagaatcgt cgtatgcagt gaaaactctc
     3781 ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga
     3841 acgacattta taatgaacgt gaattgctca acagtatggg catttcgcag cctaccgtgg
     3901 tgttcgtttc caaaaagggg ttgcaaaaaa ttttgaacgt gcaaaaaaag ctcccaatca
     3961 tccaaaaaat tattatcatg gattctaaaa cggattacca gggatttcag tcgatgtaca
     4021 cgttcgtcac atctcatcta cctcccggtt ttaatgaata cgattttgtg ccagagtcct
     4081 tcgataggga caagacaatt gcactgatca tgaactcctc tggatctact ggtctgccta
     4141 aaggtgtcgc tctgcctcat agaactgcct gcgtgagatt ctcgcatgcc agagatccta
     4201 tttttggcaa tcaaatcatt ccggatactg cgattttaag tgttgttcca ttccatcacg
     4261 gttttggaat gtttactaca ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt
     4321 atagatttga agaagagctg tttctgagga gccttcagga ttacaagatt caaagtgcgc
     4381 tgctggtgcc aaccctattc tccttcttcg ccaaaagcac tctgattgac aaatacgatt
     4441 tatctaattt acacgaaatt gcttctggtg gcgctcccct ctctaaggaa gtcggggaag
     4501 cggttgccaa gaggttccat ctgccaggta tcaggcaagg atatgggctc actgagacta
     4561 catcagctat tctgattaca cccgaggggg atgataaacc gggcgcggtc ggtaaagttg
     4621 ttccattttt tgaagcgaag gttgtggatc tggataccgg gaaaacgctg ggcgttaatc
     4681 aaagaggcga actgtgtgtg agaggtccta tgattatgtc cggttatgta aacaatccgg
     4741 aagcgaccaa cgccttgatt gacaaggatg gatggctaca ttctggagac atagcttact
     4801 gggacgaaga cgaacacttc ttcatcgttg accgcctgaa gtctctgatt aagtacaaag
     4861 gctatcaggt ggctcccgct gaattggaat ccatcttgct ccaacacccc aacatcttcg
     4921 acgcaggtgt cgcaggtctt cccgacgatg acgccggtga acttcccgcc gccgttgttg
     4981 ttttggagca cggaaagacg atgacggaaa aagagatcgt ggattacgtc gccagtcaag
     5041 taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc
     5101 ttaccggaaa actcgacgca agaaaaatca gagagatcct cataaaggcc aagaagggcg
     5161 gaaagatcgc cgtgtaattc tagagaattg tacaagtaaa gcggccgcca aagttgtttc
     5221 tgactgacta ataagtataa tttgtttcta ttatgtataa gttaagctaa ttacttattt
     5281 tataatacaa catgactgtt tttaaagtac aaaataagtt tatttttgta aaagagagaa
     5341 tgtttaaaag ttttgttact ttatagaaga aattttgagt ttttgttttt ttttaataaa
     5401 taaataaaca taaataaatt gtttgttgaa tttattatta gtatgtaagt gtaaatataa
     5461 taaaacttaa tatctattca aattaataaa taaacctcga tatacagacc gataaaacac
     5521 atgcgtcaat tttacgcatg attatcttta acgtacgtca caatatgatt atctttctag
     5581 ggttaaataa tagtttctaa tttttttatt attcagcctg ctgtcgtgaa taccgtatat
     5641 ctcaacgctg tctgtgagat tgtcgtattc tagccttttt agtttttcgc tcatcgactt
     5701 gatattgtcc gacacatttt cgtcgatttg cgttttgatc aaagacttga gcagagacac
     5761 gttaatcaac tgttcaaatt gatccatatt aacgatatca acccgatgcg tatatggtgc
     5821 gtaaaatata ttttttaacc ctcttatact ttgcactctg cgttaatacg cgttcgtggg
     5881 taccgagctc gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5941 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6001 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6061 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6121 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6181 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6241 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6301 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6361 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6421 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6481 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6541 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6601 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6661 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6721 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6781 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6841 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6901 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6961 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7021 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7081 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7141 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7201 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7261 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7321 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7381 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7441 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7501 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7561 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7621 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7681 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7741 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7801 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7861 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7921 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7981 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8041 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     8101 ataggcgtat cacgaggccc tttcgtc
//
