LOCUS       40924_5509        6062 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pBac-IE2-HsCBDBMP4-Puro DNA, complete sequence.
ACCESSION   .
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  JOURNAL   Unpublished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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6062)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-FEB-2013) Contact:Jae Man Lee Laboratory of Silkworm Science, 
            Kyushu University Graduate School of Bioresource and 
            Bioenvironmental Sciences"; volume: " 6-10-1 Hakozaki, Higashi-ku, 
            Fukuoka 812-8581, Japan E-mail :jaemanle@agr.kyushu-u.ac.jp Phone 
            :81-92-642-2842 Fax "; pages: "81-92-642-284"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6062
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        371..662
                     /label=OpIE-1 promoter
                     /note="moderate constitutive baculovirus promoter for
                     insect cell expression"
     CDS             684..1280
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    1484..1618
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1633..2180
                     /label=OpIE-2 promoter
                     /note="strong constitutive baculovirus promoter for insect
                     cell expression"
     protein_bind    2215..2235
                     /label=attB1
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     sig_peptide     2259..2321
                     /label=melittin signal sequence
                     /note="signal sequence from honeybee melittin"
     CDS             3006..3020
                     /codon_start=1
                     /label=enterokinase site
                     /note="enterokinase recognition and cleavage site"
                     /translation="DDDDK"
     CDS             3405..3425
                     /codon_start=1
                     /label=TEV site
                     /note="tobacco etch virus (TEV) protease recognition and 
                     cleavage site"
                     /translation="ENLYFQG"
     CDS             3435..3458
                     /codon_start=1
                     /label=8xHis
                     /note="8xHis affinity tag"
                     /translation="HHHHHHHH"
     protein_bind    complement(3462..3486)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3516..3557
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     CDS             3567..3584
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     polyA_signal    3602..3731
                     /label=OpIE-2 poly(A) signal
                     /note="baculovirus polyadenylation signal"
     promoter        4278..4382
                     /label=AmpR promoter
     CDS             4383..5240
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5414..6002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgataagctt gaaagtttag gtcgagtaaa gcgcaaatct tttttaaccc tagaaagata
       61 gtctgcgtaa aattgacgca tgcattcttg aaatattgct ctctctttct aaatagcgcg
      121 aatccgtcgc tgtgcattta ggacatctca gtcgccgctt ggagctcccg tgaggcgtgc
      181 ttgtcaatgc ggtaagtgtc actgattttg aactataacg accgcgtgag tcaaaatgac
      241 gcatgattat cttttacgtg acttttaaga tttaactcat acgataatta tattgttatt
      301 tcatgttcta cttacgtgat aacttattat atatatattt tcttgttata gattaagaga
      361 tcttggtcat gcgaaacacg cacggcgcgc gcacgcagct tagcacaaac gcgtcgttgc
      421 acgcgcccac cgctaaccgc aggccaatcg gtcggccggc ctcatatccg ctcaccagcc
      481 gcgtcctatc gggcgcggct tccgcgccca ttttgaataa ataaacgata acgccgttgg
      541 tggcgtgagg catgtaaaag gttacatcat tatcttgttc gccatccggt tggtataaat
      601 agacgttcat gttggttttt gtttcagttg caagttggct gcggcgcgcg cagcaccttt
      661 gccgggatct gccgggcaat tccatgaccg agtacaagcc cacggtgcgc ctcgccaccc
      721 gcgacgacgt cccccgggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca
      781 cgcgccacac cgtcgacccg gaccgccaca tcgagcgggt caccgagctg caagaactct
      841 tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg
      901 tggcggtctg gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc
      961 cgcgcatggc cgagttgagc ggttccaggc tggccgcgca gcaacagatg gaaggcctcc
     1021 tggcgccgca ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg
     1081 accaccaggg caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc
     1141 gcgccggggt gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc
     1201 ggctcggctt caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca
     1261 tgacccgcaa gcccggtgcc aattctctaa ggaaatactt aaccatggat atctaatgac
     1321 tagggataac agggtaatct agtaacagct gcccggcaac cactacgtgg actccaagct
     1381 ggacatcacc aaccacaacg aggactacac cgtggtggag cagtacgagc acgccgaggc
     1441 ccgccactcc ggctcccagt agggactctg atctcgacct cgaaacttgt ttattgcagc
     1501 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     1561 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatctc
     1621 tagactagtg ttcatgatga taaacaatgt atggtgctaa tgttgcttca acaacaattc
     1681 tgttgaactg tgttttcatg tttgccaaca agcaccttta tactcggtgg cctccccacc
     1741 accaactttt ttgcactgca aaaaaacacg cttttgcacg cgggcccata catagtacaa
     1801 actctacgtt tcgtagacta ttttacataa atagtctaca ccgttgtata cgctccaaat
     1861 acactaccac acattgaacc tttttgcagt gcaaaaaagt acgtgtcggc agtcacgtag
     1921 gccggcctta tcgggtcgcg tcctgtcacg tacgaatcac attatcggac cggacgagtg
     1981 ttgtcttatc gtgacaggac gccagcttcc tgtgttgcta accgcagccg gacgcaactc
     2041 cttatcggaa caggacgcgc ctccatatca gccgcgcgtt atctcatgcg cgtgaccgga
     2101 cacgaggcgc ccgtcccgct tatcgcgcct ataaatacag cccgcaacga tctggtaaac
     2161 acagttgaac agcatctgtt cgaatttaaa gcttggtacc gagatcgagg cctacaagtt
     2221 tgtacaaaaa agcaggcaac tcctaaaaaa ccgccaccat gaaattctta gtcaacgttg
     2281 cccttgtttt tatggtcgta tacatttctt acatctatgc ggatccagca gctgtttacc
     2341 aaccgcagcc tcacccccag cctcctccct atggccactg tgtcacagac agtggtgtgg
     2401 tctactctgt ggggatgcag tggctgaaga cacaaggaaa taagcaaatg ctttgcacgt
     2461 gcctgggcaa cggagtcagc tgccaagaga cagctgtaac ccagacttac ggtggcaact
     2521 caaatggaga gccatgtgtc ttaccattca cctacaatgg caggacgttc tactcctgca
     2581 ccacagaagg gcgacaggac ggacatcttt ggtgcagcac aacttcgaat tatgagcagg
     2641 accagaaata ctctttctgc acagaccaca ctgttttggt tcagactcga ggaggaaatt
     2701 ccaatggtgc cttgtgccac ttccccttcc tatacaacaa ccacaattac actgattgca
     2761 cttctgaggg cagaagagac aacatgaagt ggtgtgggac cacacagaac tatgatgccg
     2821 accagaagtt tgggttctgc cccatggctg cccacgagga aatctgcaca accaatgaag
     2881 gggtcatgta ccgcattgga gatcagtggg ataagcagca tgacatgggt cacatgatga
     2941 ggtgcacgtg tgttgggaat ggtcgtgggg aatggacatg cattgcctac tcgcagcttc
     3001 gagtcgacga cgatgataag cgtagcccta agcatcactc acagcgggcc aggaagaaga
     3061 ataagaactg ccggcgccac tcgctctatg tggacttcag cgatgtgggc tggaatgact
     3121 ggattgtggc cccaccaggc taccaggcct tctactgcca tggggactgc ccctttccac
     3181 tggctgacca cctcaactca accaaccatg ccattgtgca gaccctggtc aattctgtca
     3241 attccagtat ccccaaagcc tgttgtgtgc ccactgaact gagtgccatc tccatgctgt
     3301 acctggatga gtatgataag gtggtactga aaaattatca ggagatggta gtagagggat
     3361 gtgggtgccg ctctagagcg gccgccaccg cctcgagtgc aggcgaaaac ctgtacttcc
     3421 agggaagctc cagccaccac catcaccatc atcatcacta gacccagctt tcttgtacaa
     3481 agtggtgacg taagctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     3541 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt tatctgacta
     3601 aatcttagtt tgtattgtca tgttttaata caatatgtta tgtttaaata tgtttttaat
     3661 aaattttata aaataatttc aacttttatt gtaacaacat tgtccattta cacactcctt
     3721 tcaagcgcgt gaacactagt ctagagaaaa gttttgttac tttatagaag aaattttgag
     3781 tttttgtttt tttttaataa ataaataaac ataaataaat tgtttgttga atttattatt
     3841 agtatgtaag tgtaaatata ataaaactta atatctattc aaattaataa ataaacctcg
     3901 atatacagac cgataaaaca catgcgtcaa ttttacgcat gattatcttt aacgtacgtc
     3961 acaatatgat tatctttcta gggttaaata atagtttcta atttttttat tattcagcct
     4021 gctgtcgtga ataccgtata tctcaacgct gtctgtgaga ttgtcgtatt ctagcctttt
     4081 tagtttttcg ctcatcgact tgatattgtc cgacacattt tcgtcgattt gcgttttgat
     4141 caaagacttg agcagagaca cgttaatcaa ctgttcaaat tgatccatat taacgatggg
     4201 ggatcactgc ttgagcctag gaattttaac aaaatattaa cgcttacaat ttaggtggca
     4261 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4321 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4381 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4441 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4501 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4561 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4621 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4681 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4741 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4801 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4861 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     4921 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     4981 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5041 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     5101 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5161 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5221 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     5281 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     5341 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     5401 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     5461 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     5521 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
     5581 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     5641 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     5701 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     5761 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     5821 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     5881 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     5941 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6001 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     6061 tg
//
