LOCUS       Exported                8127 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  PiggyBac donor vector pPIGA3Fluc DNA, complete sequence.
ACCESSION   AB593379
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  JOURNAL   Unpublished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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8127
                     /organism="piggyBac donor vector pPIGA3Fluc"
                     /mol_type="other DNA"
                     /db_xref="taxon:905930"
     misc_feature    1..404
                     /label=cloning vector pUC18
                     /note="cloning vector pUC18"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  556..5582
                     /mobile_element_type="transposon:piggyBac"
     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"
     regulatory      1373..1765
                     /regulatory_class="promoter"
                     /gene="hsp70"
                     /note="Hsp70Bb"
     gene            1373..1765
                     /gene="hsp70"
                     /label=hsp70
     CDS             1864..2463
                     /codon_start=1
                     /gene="pac"
                     /product="puromycin N-acetyltransferase"
                     /label=pac
                     /protein_id="BAJ41850.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     gene            1864..2463
                     /gene="pac"
                     /label=pac
     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
                     /regulatory_class="promoter"
                     /gene="A3"
                     /note="actin A3"
     exon            2967..3035
                     /gene="A3"
                     /note="actin A3"
     intron          3036..3495
                     /gene="A3"
                     /note="actin A3"
     regulatory      3519..3528
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3525..5177
                     /codon_start=1
                     /gene="luc"
                     /product="luciferase"
                     /label=luc
                     /note="firefly"
                     /protein_id="BAJ41851.1"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     gene            3525..5177
                     /gene="luc"
                     /label=luc
     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"
     misc_feature    5892..8127
                     /label=cloning vector pUC18
                     /note="cloning vector pUC18"
     primer_bind     complement(5906..5922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5930..5946
                     /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)."
     promoter        complement(5954..5984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5999..6020
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /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(7067..7927)
                     /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"
     promoter        complement(7928..8032)
                     /gene="bla"
                     /label=AmpR promoter