LOCUS       Exported                9375 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  PiggyBac donor vector pPIGA3 DNA, complete sequence.
ACCESSION   AB593377
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9375
                     /organism="piggyBac donor vector pPIGA3"
                     /mol_type="other DNA"
                     /db_xref="taxon:905928"
     misc_feature    1..2241
                     /label=cloning vector pUC18
                     /note="cloning vector pUC18"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             201..1061
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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
                     /bound_moiety="lac repressor encoded by lacI"
                     /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"
     mobile_element  2375..8292
                     /mobile_element_type="transposon:piggyBac"
     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"
     regulatory      3191..3583
                     /regulatory_class="promoter"
                     /gene="hsp70"
                     /note="Hsp70Bb"
     gene            3191..3583
                     /gene="hsp70"
                     /label=hsp70
     CDS             3682..4281
                     /codon_start=1
                     /gene="pac"
                     /product="puromycin N-acetyltransferase"
                     /label=pac
                     /protein_id="BAJ41846.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     gene            3682..4281
                     /gene="pac"
                     /label=pac
     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
                     /regulatory_class="promoter"
                     /gene="A3"
                     /note="actin A3"
     exon            4785..4853
                     /gene="A3"
                     /note="actin A3"
     intron          4854..5313
                     /gene="A3"
                     /note="actin A3"
     regulatory      5337..5346
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5343..6062
                     /codon_start=1
                     /gene="EGFP"
                     /product="enhanced green fluorescent protein"
                     /label=EGFP
                     /protein_id="BAJ41847.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            5343..6062
                     /gene="EGFP"
                     /label=EGFP
     misc_feature    6079..6305
                     /gene="SV40"
                     /label=SV40 polyadenylation sequence
                     /note="SV40 polyadenylation sequence"
     gene            6079..6305
                     /gene="SV40"
                     /label=SV40
     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"
     misc_feature    8972..9375
                     /label=cloning vector pUC18
                     /note="cloning vector pUC18"
     primer_bind     complement(8981..8997)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"