LOCUS       Exported               14183 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pUB-GWS-Hyg DNA, complete sequence.
ACCESSION   AB303067
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14183)
  AUTHORS   Maekawa T, Kusakabe M, Shimoda Y, Sato S, Tabata S, Murooka Y, 
            Hayashi M.
  TITLE     Polyubiquitin Promoter-Based Binary Vectors for Overexpression and 
            Gene Silencing in Lotus japonicus
  JOURNAL   Mol. Plant Microbe Interact. 21 (4), 375-382 (2008)
  PUBMED    18321183
REFERENCE   2  (bases 1 to 14183)
  AUTHORS   Maekawa T, Hayashi M.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAY-2007) Contact:Makoto Hayashi National Institute of
            Agrobiological Sciences, Environmental Stress Research Unit; 2-1-2 
            Kannondai, Tsukuba, Ibaraki 305-8602, Japan
REFERENCE   3  (bases 1 to 14183)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14183
                     /organism="Binary vector pUB-GWS-Hyg"
                     /mol_type="other DNA"
                     /clone="pUB-GWS-Hyg"
                     /db_xref="taxon:442629"
     misc_feature    1..26
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(93..301)
                     /label=CaMV 3 UTR (polyA signal)
                     /note="CaMV 3 UTR (polyA signal)"
     polyA_signal    103..277
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(317..1342)
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin phosphotransferase"
                     /label=hptII
                     /note="hptII (hygromycin resistance) gene"
                     /protein_id="BAG13008.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(317..1342)
                     /gene="hptII"
                     /label=hptII
     regulatory      complement(1378..2158)
                     /regulatory_class="promoter"
                     /note="CaMV35S; 35S promoter from CaMV"
     promoter        complement(1410..2087)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    2278..2299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2314..2344
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2352..2368
                     /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)."
     primer_bind     2376..2392
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(2414..2667)
                     /label=Nos terminator amplified from pBI121
                     /note="Nos terminator amplified from pBI121"
     terminator      2414..2666
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2743..2867
                     /label=attR1 site
                     /note="attR1 site"
     protein_bind    2743..2867
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2892..2922
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2976..3635
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="cat (Chloramphenicol resistance) gene"
                     /protein_id="BAG13009.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            2976..3635
                     /gene="cat"
                     /label=cat
     CDS             3977..4282
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BAG13010.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            3977..4282
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    4323..4447
                     /label=attR2 site
                     /note="attR2 site"
     protein_bind    complement(4323..4447)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     misc_feature    4481..4735
                     /note="contains intron 1 of Arabidopsis thaliana WRKY 
                     transcription factor 33"
     misc_feature    complement(4790..4914)
                     /label=attR2 site
                     /note="attR2 site"
     protein_bind    4790..4914
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4955..5260)
                     /codon_start=1
                     /gene="ccdB"
                     /label=ccdB
                     /protein_id="BAG13011.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            complement(4955..5260)
                     /gene="ccdB"
                     /label=ccdB
     CDS             complement(5602..6261)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /note="cat (Chloramphenicol resistance) gene"
                     /protein_id="BAG13012.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(5602..6261)
                     /gene="cat"
                     /label=cat
     promoter        complement(6315..6345)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     misc_feature    complement(6370..6494)
                     /label=attR1 site
                     /note="attR1 site"
     protein_bind    complement(6370..6494)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(6522..6538)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     gene            complement(6539..7125)
                     /gene="Ljubq1"
                     /label=Ljubq1
     intron          complement(6539..7078)
                     /gene="Ljubq1"
                     /note="5 UTR intron of Ljubq1 gene"
     exon            complement(7079..7125)
                     /gene="Ljubq1"
                     /number=1
     regulatory      complement(7126..7660)
                     /regulatory_class="promoter"
                     /note="Ljubq1 promoter"
     primer_bind     complement(7687..7703)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7906..7930
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    7925..7950
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"
     CDS             9230..9859
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             10293..11360
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      complement(10584..11584)
                     /direction=LEFT
                     /note="pVS1-REP; replication origin from pVS1"
     rep_origin      11426..11620
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    11964..12104
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(11994..12254)
                     /label=bom site from pBR322
                     /note="bom site from pBR322"
     rep_origin      complement(12290..12878)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(12394..12674)
                     /direction=LEFT
                     /label=pBR322 origin of replication
                     /note="pBR322 origin of replication"
     CDS             complement(12965..13759)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside phosphotransferase"
                     /label=aadA
                     /note="aadA (kanamycin resistance) gene amplified from 
                     pIG121Hm"
                     /protein_id="BAG13013.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(12965..13759)
                     /gene="aadA"
                     /label=aadA