LOCUS       Exported               12072 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pINDEX4, complete sequence.
ACCESSION   AF294981
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12072)
  AUTHORS   Ouwerkerk PB, de Kam RJ, Hoge JH, Meijer AH.
  TITLE     Glucocorticoid-inducible gene expression in rice
  JOURNAL   Planta 213 (3), 370-378 (2001)
  PUBMED    11506359
REFERENCE   2  (bases 1 to 12072)
  AUTHORS   Ouwerkerk PBF.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-AUG-2000) Dept. of Molecular Cell Biology, Institute 
            for Molecular Plant Sciences, Wassenaarseweg 64, Leiden 2333 AL, 
            Netherlands
REFERENCE   3  (bases 1 to 12072)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12072
                     /organism="Binary vector pINDEX4"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:142848"
     misc_feature    1..12072
                     /note="pINDEX4 is a binary vector designed for 
                     glucocorticoid-inducible gene expression in plants and is 
                     optimized for use in rice; pINDEX4 is part of a series of 
                     four pINDEX vectors (Genbank Accession Numbers 
                     AF294979-AF294982). pINDEX vectors are based on parts of 
                     pCAMBIA-1300 encoded by GenBank Accession Number AF234296 
                     and pTA7002 (Aoyama, T. and Chua N.-H., 1997, Plant J. 
                     11:605-612)"
     misc_feature    36..74
                     /note="multiple cloning site; restriction sites 
                     HpaI-ApaI-MunI-MluI-SwaI are unique. In here a promoter of 
                     choice to drive GVG expression can be inserted."
     gene            81..1394
                     /gene="GVG"
                     /label=GVG
     CDS             93..1394
                     /codon_start=1
                     /gene="GVG"
                     /product="GVG"
                     /label=GVG
                     /note="chimeric transcription factor; contains 
                     Saccharomyces cerevisiae GAL4BD, Herpes simplex VP16AD and 
                     Rattus norvegicus GR domain"
                     /protein_id="AAG38033.1"
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPRSSAPPTDVSLGDELHLDGEDVAMAHADALD
                     DFDLDMLGDGDSPGPGFTPHDSAPYGALDMADFEFEQMFTDALGIDEYGGDPIQQATAG
                     VSQDTSENPNKTIVPAALPQLTPTLVSLLEVIEPEVLYAGYDSSVPDSAWRIMTTLNML
                     GGRQVIAAVKWAKAIPGFRNLHLDDQMTLLQYSWMFLMAFALGWRSYRQSSGNLLCFAP
                     DLIINEQRMSLPCMYDQCKHMLFVSSELQRLQVSYEEYLCMKTLLLLSSVPKEGLKSQE
                     LFDEIRMTYIKELGKAIVKREGNSSQNWQRFYQLTKLLDSMHEVVENLLTYCFQTFLDK
                     TMSIEFPEMLAEIITNQIPKYSNGNIKKLLFHQK"
     CDS             321..554
                     /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"
     3UTR           1421..1717
                     /label=Pisum sativum RbcS-E9 polyA signal
                     /note="Pisum sativum RbcS-E9 polyA signal"
     terminator      1895..2147
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      2199..2269
                     /regulatory_class="promoter"
                     /note="4UAS; tetramer of the GVG binding site"
     regulatory      2316..2373
                     /regulatory_class="promoter"
                     /note="CaMV 35S"
     promoter        2318..2364
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    2395..2452
                     /note="multiple cloning site; restriction sites 
                     BbrPI-SpeI-Sse8387I-XhoI-Bsp1407I-StuI are unique."
     3UTR           2456..2924
                     /label=Pisum sativum RbcS-3A polyA signal
                     /note="Pisum sativum RbcS-3A polyA signal"
     primer_bind     complement(2963..2979)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3182..3206
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    3201..3226
                     /label=right border repeat from C58 T-DNA
                     /note="right border repeat from C58 T-DNA"
     CDS             4506..5135
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5569..6636
                     /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      6702..6896
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    7240..7380
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7566..8154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8241..9035)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    9460..9485
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    9460..9484
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     3UTR           9552..9771
                     /label=CaMV (polyA signal
                     /note="CaMV (polyA signal"
     polyA_signal    9562..9736
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(join(9787..10479,10670..11002))
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin phosphotransferase"
                     /label=hptII
                     /note="hygromycin resistance"
                     /protein_id="AAG38032.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(9787..11002)
                     /gene="hptII"
                     /label=hptII
     exon            complement(9787..10439)
                     /gene="hptII"
                     /number=2
     intron          10474..10663
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     intron          complement(10480..10669)
                     /gene="hptII"
                     /note="derived from Ricinus communis catalase-1 intron 
                     presented in Genbank Accession Number D21161"
     exon            complement(10670..11002)
                     /gene="hptII"
                     /number=1
     regulatory      complement(11049..11829)
                     /regulatory_class="promoter"
                     /note="CaMV35S; 35S promoter from CaMV"
     promoter        complement(11081..11758)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    11949..11970
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11985..12015
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    12023..12039
                     /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     12047..12063
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"