LOCUS       Exported                6062 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pBac-IE2-HsCBDBMP4-Puro DNA, complete sequence.
ACCESSION   AB794658
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6062)
  AUTHORS   Imai S, Li Z, Miyagawa Y, Toyoda M, Umezawa A, Mon H, Kusakabe T, 
            Yoshimura K, Lee JM.
  TITLE     Biologically active human bone morphogenetic protein 4-fused to 
            collagen binding domain produced in silkworm-baculovirus expression 
            system
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6062)
  AUTHORS   Lee JM, Li Z, Kusakabe T.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-FEB-2013) Contact:Jae Man Lee Laboratory of Silkworm 
            Science, Kyushu University Graduate School of Bioresource and 
            Bioenvironmental Sciences; 6-10-1 Hakozaki, Higashi-ku, Fukuoka 
            812-8581, Japan E-mail :jaemanle@agr.kyushu-u.ac.jp Phone 
            :81-92-642-2842 Fax :81-92-642-2842
REFERENCE   3  (bases 1 to 6062)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6062
                     /organism="Expression vector pBac-IE2-HsCBDBMP4-Puro"
                     /mol_type="other DNA"
                     /db_xref="taxon:1298532"
     promoter        371..662
                     /gene="immediate-early gene 1 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-1 promoter
                     /note="moderate constitutive baculovirus promoter for 
                     insect cell expression"
     CDS             684..1316
                     /codon_start=1
                     /gene="Puro"
                     /product="puromycin resistance protein"
                     /label=Puro
                     /protein_id="BAM95187.1"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGANSLRKYLTMDI"
     gene            684..1316
                     /gene="Puro"
                     /label=Puro
     CDS             684..1280
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    1484..1618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1633..2180
                     /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    2214..2238
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2259..3461
                     /codon_start=1
                     /gene="HsCBDBMP4"
                     /product="fusion proteins of collagen binding domain and 
                     bone morphogenetic protein 4 protein"
                     /label=HsCBDBMP4
                     /protein_id="BAM95188.1"
                     /translation="MKFLVNVALVFMVVYISYIYADPAAVYQPQPHPQPPPYGHCVTDS
                     GVVYSVGMQWLKTQGNKQMLCTCLGNGVSCQETAVTQTYGGNSNGEPCVLPFTYNGRTF
                     YSCTTEGRQDGHLWCSTTSNYEQDQKYSFCTDHTVLVQTRGGNSNGALCHFPFLYNNHN
                     YTDCTSEGRRDNMKWCGTTQNYDADQKFGFCPMAAHEEICTTNEGVMYRIGDQWDKQHD
                     MGHMMRCTCVGNGRGEWTCIAYSQLRVDDDDKRSPKHHSQRARKKNKNCRRHSLYVDFS
                     DVGWNDWIVAPPGYQAFYCHGDCPFPLADHLNSTNHAIVQTLVNSVNSSIPKACCVPTE
                     LSAISMLYLDEYDKVVLKNYQEMVVEGCGCRSRAAATASSAGENLYFQGSSSHHHHHHH
                     H"
     gene            2259..3461
                     /gene="HsCBDBMP4"
                     /label=HsCBDBMP4
     CDS             2259..2321
                     /codon_start=1
                     /product="signal sequence from honeybee melittin"
                     /label=melittin signal sequence
                     /translation="MKFLVNVALVFMVVYISYIYA"
     CDS             3006..3020
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
     CDS             3405..3425
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             3435..3458
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     protein_bind    complement(3462..3486)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3516..3557
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3567..3584
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3602..3731
                     /gene="immediate-early gene 2 from the Orgyia pseudotsugata
                     multicapsid nucleopolyhedrovirus"
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     promoter        4278..4382
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4383..5243
                     /codon_start=1
                     /gene="Amp"
                     /product="ampicillin resistance protein"
                     /label=Amp
                     /protein_id="BAM95189.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4383..5243
                     /gene="Amp"
                     /label=Amp
     rep_origin      5388..6061
                     /label=pUC replication origin
                     /note="pUC replication origin"
     rep_origin      5414..6002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"