LOCUS       Exported                7468 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pEAK4IAC-EGFP DNA, complete sequence.
ACCESSION   AB830532
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7468)
  AUTHORS   Xu J, Yoshimura K, Mon H, Li Z, Zhu L, Iiyama K, Kusakabe T, Lee JM.
  TITLE     Establishment of Caenorhabditis elegans SID-1-dependent DNA delivery
            system in cultured silkworm cells
  JOURNAL   Mol. Biotechnol. 56 (3), 193-198 (2014)
  PUBMED    23979877
REFERENCE   2  (bases 1 to 7468)
  AUTHORS   Yoshimura K, Tatsuke T, Xu J, Mon H, Lee JM, Kusakabe T.
  TITLE     Establishment of an EBNA1/oriP-based episomal vector system in 
            cultured insect cells
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 7468)
  AUTHORS   Yoshimura K, Xu J, Lee JM, Kusakabe T.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JUL-2013) Contact:Takahiro Kusakabe Kyushu university,
            Graduate School of Bioresource and Bioenvironmental Sciences, Kyushu
            University, Laboratory of Silkworm Science; 6-10-1 Hakozaki, 
            Higashi-ku, Fukuoka, Fukuoka 812-8581, Japan, Fukuoka 812-8581, 
            Japan
REFERENCE   4  (bases 1 to 7468)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7468
                     /organism="Expression vector pEAK4IAC-EGFP"
                     /mol_type="other DNA"
                     /db_xref="taxon:1355155"
     regulatory      10..558
                     /regulatory_class="promoter"
                     /gene="IE2"
                     /note="IE2 promoter"
     gene            10..558
                     /gene="IE2"
                     /label=IE2
     promoter        10..557
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect 
                     cell expression"
     protein_bind    656..680
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             722..1453
                     /codon_start=1
                     /gene="EGFP"
                     /product="enhanced green fluorescent protein"
                     /label=EGFP
                     /protein_id="BAN62844.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKHRST"
     gene            722..1453
                     /gene="EGFP"
                     /label=EGFP
     CDS             722..1438
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     protein_bind    complement(1468..1492)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1527..1568
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1578..1595
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1613..1742
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     polyA_signal    1750..1884
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1912..2511)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     promoter        complement(2533..2824)
                     /gene="immediate-early gene 1 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-1 promoter
                     /note="moderate constitutive baculovirus promoter for 
                     insect cell expression"
     polyA_signal    2854..2988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     regulatory      4250..4971
                     /regulatory_class="promoter"
                     /gene="A3"
                     /note="A3 promoter"
     gene            4250..4971
                     /gene="A3"
                     /label=A3
     rep_origin      5622..5757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6440..7300
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDHLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     terminator      7298..7433
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"