LOCUS       Exported               14533 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Gateway binary vector pGWB501_TOPX DNA, complete sequence.
ACCESSION   LC217876
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14533)
  AUTHORS   Hanano S, Ozawa K, Nagase T, Suzuki H, Sakurai N, Shibata D.
  TITLE     Construction of the Gateway cloning compatible binary vector 
            pGWB501_TOPX for induction of a foreign gene in plant
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 14533)
  AUTHORS   Hanano S, Shibata D.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-FEB-2017) Contact:Shigeru Hanano Kazusa DNA Research 
            Institute, Metabolomics; Kazusa-kamatari, 2-6-7, Kisarazu, Chiba 
            292-0818, Japan URL :http://www.kazusa.or.jp
REFERENCE   3  (bases 1 to 14533)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14533
                     /organism="Gateway binary vector pGWB501_TOPX"
                     /mol_type="other DNA"
                     /clone="pGWB501_TOPX"
                     /db_xref="taxon:1968272"
     misc_feature    1..303
                     /label=Gateway binary vector pGWB501
                     /note="Gateway binary vector pGWB501"
     misc_feature    54..78
                     /label=T-DNA right border
                     /note="T-DNA right border"
     primer_bind     280..297
                     /label=M13-fwd
                     /note="M13-fwd"
     primer_bind     281..297
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    307..577
                     /label=LexA operator
                     /note="LexA operator"
     promoter        595..641
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    599..657
                     /label=-46 to +12 of 35S promoter
                     /note="-46 to +12 of 35S promoter"
     misc_feature    661..2431
                     /label=Gateway binary vector pGWB501
                     /note="Gateway binary vector pGWB501"
     misc_feature    693..817
                     /label=attR1
                     /note="attR1"
     protein_bind    695..819
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        856..886
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             940..1599
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="BBA21867.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            940..1599
                     /gene="cat"
                     /label=cat
     CDS             1941..2246
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BBA21868.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            1941..2246
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    2287..2411
                     /label=attR2
                     /note="attR2"
     protein_bind    complement(2287..2411)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      2433..2682
                     /regulatory_class="terminator"
                     /note="HSP18.2 terminator"
     misc_feature    2688..3109
                     /label=Gateway binary vector pGWB501
                     /note="Gateway binary vector pGWB501"
     terminator      2699..2951
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(2984..3000)
                     /label=M13-rv
                     /note="M13-rv"
     protein_bind    3008..3024
                     /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(3032..3062)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3077..3098
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      3117..3864
                     /regulatory_class="promoter"
                     /note="35S promoter"
     promoter        3192..3868
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     regulatory      3876..3942
                     /regulatory_class="enhancer"
                     /note="omega translational enhancer"
     CDS             3978..5429
                     /codon_start=1
                     /gene="XVE"
                     /label=XVE
                     /protein_id="BBA21869.1"
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAAGEPSSAPPTDVSLGDELHLD
                     GEDVAMAHADALDDFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGID
                     EYGGDPSAGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPT
                     RPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVW
                     RSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSII
                     LLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLIL
                     SHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLAT
                     AGSTSSHSLQKYYITGEAEGFPATV"
     gene            3978..5429
                     /gene="XVE"
                     /label=XVE
     CDS             3978..4229
                     /codon_start=1
                     /gene="lexA (truncated)"
                     /product="DNA-binding domain of E. coli LexA"
                     /label=LexA BD
                     /note="binds the 16-bp palindromic sequence 
                     5-CTGTATATATATACAG-3"
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAA"
     CDS             4245..4478
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             4536..5429
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     regulatory      5444..5537
                     /regulatory_class="terminator"
                     /note="rat glucocorticoid receptor 3UTR"
     regulatory      5563..5858
                     /regulatory_class="terminator"
                     /note="pea rbcS E9 terminator"
     misc_feature    5866..14533
                     /label=Gateway binary vector pGWB501
                     /note="Gateway binary vector pGWB501"
     terminator      5890..6142
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      complement(5892..6157)
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     CDS             complement(6293..7318)
                     /codon_start=1
                     /gene="hpt"
                     /product="hygromycin phosphotransferase"
                     /label=hpt
                     /protein_id="BBA21870.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGAFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     gene            complement(6293..7318)
                     /gene="hpt"
                     /label=hpt
     regulatory      complement(7346..7632)
                     /regulatory_class="promoter"
                     /note="NOS promoter"
     promoter        complement(7370..7549)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    complement(8055..8079)
                     /label=T-DNA left border
                     /note="T-DNA left border"
     CDS             8600..9391
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      9637..10225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    10411..10551
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      10895..11089
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(11155..12228)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     CDS             complement(12657..13286)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"