LOCUS       Exported                4989 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pTD1-phiC31Int2 DNA, complete sequence.
ACCESSION   AB713997
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)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4989
                     /organism="Expression vector pTD1-phiC31Int2"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /db_xref="taxon:1177636"
     misc_feature    join(1..307,2276..4989)
                     /label=pTD1(AB194742)-derived sequence
                     /note="pTD1(AB194742)-derived sequence"
     regulatory      240..258
                     /regulatory_class="promoter"
                     /note="T7 RNA polymerase promoter"
     promoter        240..258
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5UTR           259..304
                     /label=Malacosma neustria NPV polyhedrin gene 5 UTR
                     /note="Malacosma neustria NPV polyhedrin gene 5 UTR"
     misc_feature    308..342
                     /label=pXINSECT DEST39-derived sequence
                     /note="pXINSECT DEST39-derived sequence"
     misc_recomb     323..347
                     /label=lambda phage attB1 sequence
                     /note="lambda phage attB1 sequence"
     protein_bind    323..347
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    328..404
                     /label=pENTR11-derived sequence
                     /note="pENTR11-derived sequence"
     misc_feature    387..392
                     /label=NcoI site was replaced with CCGTGG
                     /note="NcoI site was replaced with CCGTGG"
     CDS             407..2224
                     /codon_start=1
                     /gene="int"
                     /product="phiC31 Integrase"
                     /function="site-specific recombinase"
                     /label=int
                     /note="integrase gene of Streptomyces phage phiC31"
                     /protein_id="BAM36534.1"
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     gene            407..2224
                     /gene="int"
                     /label=int
     misc_feature    2224..2251
                     /label=pENTR11-derived sequence
                     /note="pENTR11-derived sequence"
     misc_recomb     complement(2230..2254)
                     /label=lambda phage attB2 sequence
                     /note="lambda phage attB2 sequence"
     protein_bind    complement(2230..2254)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2233..2280
                     /label=pXINSECT DEST39-derived sequence
                     /note="pXINSECT DEST39-derived sequence"
     3UTR           2295..2668
                     /label=Autographa californica NPV polyhedrin gene 3 UTR
                     /note="Autographa californica NPV polyhedrin gene 3 UTR"
     3UTR           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"
     regulatory      2695..2742
                     /regulatory_class="terminator"
                     /note="T7 RNA polymerase terminator"
     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    2792..2808
                     /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(2816..2846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2861..2882
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3152..3834)
                     /direction=LEFT
     rep_origin      complement(3170..3758)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3929..4789)
                     /codon_start=1
                     /gene="AmpR"
                     /product="beta-lactamase"
                     /label=AmpR
                     /protein_id="BAM36535.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3929..4789)
                     /gene="AmpR"
                     /label=AmpR
     promoter        complement(4790..4894)
                     /gene="bla"
                     /label=AmpR promoter