LOCUS       40924_42944        4989 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTD1-phiC31Int2 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4989)
  AUTHORS   Yonemura N, Tamura T, Uchino K, Kobayashi I, Tatematsu K, Iizuka T, 
            Sezutsu H, Muthulakshmi M, Nagaraju J, Kusakabe T.
  TITLE     PhiC31 integrase-mediated cassette exchange in silkworm embryos
  JOURNAL   Mol. Genet. Genomics 287 (9), 731-739 (2012)
  PUBMED    22842670
REFERENCE   2  (bases 1 to 4989)
  AUTHORS   Yonemura N, Kobayashi I, Uchino K, Sezutsu H, Tamura T, Kusakabe T.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-APR-2012) Contact:Naoyuki Yonemura National Institute 
            of Agrobiological Sciences (NIAS), MAFF; Ohwashi 1-2, Tsukuba 
            Science City 305-8634, Japan
REFERENCE   3  (bases 1 to 4989)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4989)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Genet.
            Genomics"; date: "2012"; volume: "287"; issue: "9"; pages: "731-739"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (21-APR-2012) Contact:Naoyuki Yonemura National Institute of 
            Agrobiological Sciences (NIAS), MAFF; Ohwashi 1-2, Tsukuba Science 
            City 305-8634, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..4989
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        240..258
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           259..304
                     /label=polyhedrin 5' UTR
                     /note="5' UTR from the Malacosoma neustria
                     nucleopolyhedrovirus polyhedrin gene (Ezure et al., 2006)"
     protein_bind    323..347
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    387..392
                     /label=NcoI site was replaced with CCGTGG
                     /note="NcoI site was replaced with CCGTGG"
     CDS             407..2221
                     /codon_start=1
                     /label=phage phi-C31 integrase
                     /note="integrase from phage phi-C31"
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     protein_bind    complement(2230..2254)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     3'UTR           2296..2668
                     /label=polyhedrin 3' UTR
                     /note="3' UTR from the Autographa californica 
                     nucleopolyhedrovirus polyhedrin gene (Suzuki et al., 2006)"
     misc_feature    2673..2694
                     /label=synthetic poly(A)20 region
                     /note="synthetic poly(A)20 region"
     terminator      2695..2742
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2768..2784)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2792..2808)
                     /label=lac operator
                     /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(2816..2846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(2861..2882)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3170..3758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3932..4789)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4790..4894)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcctt
      241 aatacgactc actataggag tatttttatt ctttcgtaaa aaaattagaa aaataaaata
      301 taaagatgct ggatcctcga tcacaagttt gtacaaaaaa gcaggcttcg aaggagatag
      361 aaccaattct ctaaggaaat acttaaccgt ggtcgactgg atccggatgg acacgtacgc
      421 gggtgcttac gaccgtcagt cgcgcgagcg cgagaattcg agcgcagcaa gcccagcgac
      481 acagcgtagc gccaacgaag acaaggcggc cgaccttcag cgcgaagtcg agcgcgacgg
      541 gggccggttc aggttcgtcg ggcatttcag cgaagcgccg ggcacgtcgg cgttcgggac
      601 ggcggagcgc ccggagttcg aacgcatcct gaacgaatgc cgcgccgggc ggctcaacat
      661 gatcattgtc tatgacgtgt cgcgcttctc gcgcctgaag gtcatggacg cgattccgat
      721 tgtctcggaa ttgctcgccc tgggcgtgac gattgtttcc actcaggaag gcgtcttccg
      781 gcagggaaac gtcatggacc tgattcacct gattatgcgg ctcgacgcgt cgcacaaaga
      841 atcttcgctg aagtcggcga agattctcga cacgaagaac cttcagcgcg aattgggcgg
      901 gtacgtcggc gggaaggcgc cttacggctt cgagcttgtt tcggagacga aggagatcac
      961 gcgcaacggc cgaatggtca atgtcgtcat caacaagctt gcgcactcga ccactcccct
     1021 taccggaccc ttcgagttcg agcccgacgt aatccggtgg tggtggcgtg agatcaagac
     1081 gcacaaacac cttcccttca agccgggcag tcaagccgcc attcacccgg gcagcatcac
     1141 ggggctttgt aagcgcatgg acgctgacgc cgtgccgacc cggggcgaga cgattgggaa
     1201 gaagaccgct tcaagcgcct gggacccggc aaccgttatg cgaatccttc gggacccgcg
     1261 tattgcgggc ttcgccgctg aggtgatcta caagaagaag ccggacggca cgccgaccac
     1321 gaagattgag ggttaccgca ttcagcgcga cccgatcacg ctccggccgg tcgagcttga
     1381 ttgcggaccg atcatcgagc ccgctgagtg gtatgagctt caggcgtggt tggacggcag
     1441 ggggcgcggc aaggggcttt cccgggggca agccattctg tccgccatgg acaagctgta
     1501 ctgcgagtgt ggcgccgtca tgacttcgaa gcgcggggaa gaatcgatca aggactctta
     1561 ccgctgccgt cgccggaagg tggtcgaccc gtccgcacct gggcagcacg aaggcacgtg
     1621 caacgtcagc atggcggcac tcgacaagtt cgttgcggaa cgcatcttca acaagatcag
     1681 gcacgccgaa ggcgacgaag agacgttggc gcttctgtgg gaagccgccc gacgcttcgg
     1741 caagctcact gaggcgcctg agaagagcgg cgaacgggcg aaccttgttg cggagcgcgc
     1801 cgacgccctg aacgcccttg aagagctgta cgaagaccgc gcggcaggcg cgtacgacgg
     1861 acccgttggc aggaagcact tccggaagca acaggcagcg ctgacgctcc ggcagcaagg
     1921 ggcggaagag cggcttgccg aacttgaagc cgccgaagcc ccgaagcttc cccttgacca
     1981 atggttcccc gaagacgccg acgctgaccc gaccggccct aagtcgtggt gggggcgcgc
     2041 gtcagtagac gacaagcgcg tgttcgtcgg gctcttcgta gacaagatcg ttgtcacgaa
     2101 gtcgactacg ggcagggggc agggaacgcc catcgagaag cgcgcttcga tcacgtgggc
     2161 gaagccgccg accgacgacg acgaagacga cgcccaggac ggcacggaag acgtagcggc
     2221 gtaatctaga cccagctttc ttgtacaaag tggtgataat taattaagat cgaatctaga
     2281 gtcgggcggc tgtaaaacac gatacattgt tattagtaca tttattaagc gctagattct
     2341 gtgcgttgtt gatttacaga caattgttgt acgtatttta ataattcatt aaatttataa
     2401 tctttagggt ggtatgttag agcgaaaatc aaatgatttt cagcgtcttt atatctgaat
     2461 ttaaatatta aatcctcaat agatttgtaa aataggtttc gattagtttc aaacaagggt
     2521 tgtttttccg aaccgatggc tggactatct aatggatttt cgctcaacgc cacaaaactt
     2581 gccaaatctt gtagcagcaa tctagctttg tcgatattcg tttgtgtttt gttttgtaat
     2641 aaaggttcga cgtcgttcaa aatattatcc gcaaaaaaaa aaaaaaaaaa aaaactagca
     2701 taaccccttg gggcctctaa acgggtcttg aggggttttt tgcggccgca agcttggcgt
     2761 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     2821 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     2881 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     2941 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     3001 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     3061 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     3121 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3181 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     3241 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     3301 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     3361 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     3421 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     3481 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     3541 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     3601 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     3661 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     3721 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     3781 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     3841 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     3901 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     3961 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4021 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     4081 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4141 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4201 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     4261 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     4321 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     4381 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     4441 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     4501 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     4561 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     4621 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     4681 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     4741 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     4801 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     4861 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     4921 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     4981 cctttcgtc
//
