LOCUS       40924_34629        7172 bp DNA     circular SYN 18-DEC-2018
DEFINITION  PiggyBac donor vector pPIGA3II DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7172)
  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  JOURNAL   Unpublished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 7172)
  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 7172)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7172)
  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            "Unpublished"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..7172
                     /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..4241
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    4440..4463
                     /note="piggyBac transposase polyadenylation sequence; 
                     transposase"
     repeat_region   4565..4627
                     /label=piggyBac 3' inverted terminal repeat
                     /note="piggyBac 3' inverted terminal repeat"
     primer_bind     complement(4951..4967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4975..4991)
                     /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(4999..5029)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5044..5065)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(5353..5941)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6115..6972)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6973..7077)
                     /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 caccatggtg agcaagggcg
     3541 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     3601 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     3661 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     3721 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     3781 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     3841 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     3901 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     3961 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     4021 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     4081 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     4141 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     4201 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgccaaagtt
     4261 gtttctgact gactaataag tataatttgt ttctattatg tataagttaa gctaattact
     4321 tattttataa tacaacatga ctgtttttaa agtacaaaat aagtttattt ttgtaaaaga
     4381 gagaatgttt aaaagttttg ttactttata gaagaaattt tgagtttttg ttttttttta
     4441 ataaataaat aaacataaat aaattgtttg ttgaatttat tattagtatg taagtgtaaa
     4501 tataataaaa cttaatatct attcaaatta ataaataaac ctcgatatac agaccgataa
     4561 aacacatgcg tcaattttac gcatgattat ctttaacgta cgtcacaata tgattatctt
     4621 tctagggtta aataatagtt tctaattttt ttattattca gcctgctgtc gtgaataccg
     4681 tatatctcaa cgctgtctgt gagattgtcg tattctagcc tttttagttt ttcgctcatc
     4741 gacttgatat tgtccgacac attttcgtcg atttgcgttt tgatcaaaga cttgagcaga
     4801 gacacgttaa tcaactgttc aaattgatcc atattaacga tatcaacccg atgcgtatat
     4861 ggtgcgtaaa atatattttt taaccctctt atactttgca ctctgcgtta atacgcgttc
     4921 gtgggtaccg agctcgaatt cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     4981 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     5041 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     5101 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     5161 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     5221 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     5281 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5341 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5401 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5461 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5521 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5581 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5641 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5701 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5761 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5821 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5881 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5941 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     6001 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     6061 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     6121 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     6181 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     6241 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     6301 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     6361 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     6421 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     6481 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6541 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6601 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6661 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6721 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6781 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6841 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6901 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6961 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     7021 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     7081 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     7141 taaaaatagg cgtatcacga ggccctttcg tc
//
