LOCUS       Exported               11801 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pJZ254, complete sequence.
ACCESSION   KP244680
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11801)
  AUTHORS   Zhang J, Khan SA, Hasse C, Ruf S, Heckel DG, Bock R.
  TITLE     Pest control. Full crop protection from an insect pest by expression
            of long double-stranded RNAs in plastids
  JOURNAL   Science 347 (6225), 991-994 (2015)
  PUBMED    25722411
REFERENCE   2  (bases 1 to 11801)
  AUTHORS   Zhang J, Khan SA, Hasse C, Ruf S, Heckel DG, Bock R.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-DEC-2014) Bock, Max Planck Institute of Molecular 
            Plant Physiology, Am Muehlenberg1, potsdam, Brandenburg 14476, 
            Germany
REFERENCE   3  (bases 1 to 11801)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11801
                     /organism="Binary vector pJZ254"
                     /mol_type="other DNA"
                     /db_xref="taxon:1622251"
     promoter        21..39
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      69..1404
                     /regulatory_class="promoter"
                     /gene="dual CaMV 35S"
     gene            69..1404
                     /gene="dual CaMV 35S"
                     /label=dual CaMV 35S
     promoter        1057..1402
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    1437..1656
                     /label=similar to Leptinotarsa decemlineata shrub gene
                     /note="similar to Leptinotarsa decemlineata shrub gene"
     intron          1668..1868
                     /gene="St-GA20 ox"
                     /number=1
                     /note="first intron from potato GA20 oxidase gene"
     gene            1668..1868
                     /gene="St-GA20 ox"
                     /label=St-GA20 ox
                     /note="GA20 oxidase"
     misc_feature    complement(1875..2094)
                     /label=similar to Leptinotarsa decemlineata shrub gene
                     /note="similar to Leptinotarsa decemlineata shrub gene"
     terminator      2104..2811
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     regulatory      2104..2810
                     /regulatory_class="terminator"
                     /note="octopine synthase gene terminator"
     promoter        complement(2852..2870)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2903..2919)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   3121..3144
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    3122..3146
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4206..5206
                     /label=pVS1
                     /note="pVS1"
     CDS             4446..5075
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5509..6576
                     /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      6642..6836
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    7180..7320
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7506..8094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      7609..7890
                     /label=pBR325
                     /note="pBR325"
     CDS             complement(8181..8975)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /function="kanamycin resistance"
                     /label=aphA-3
                     /protein_id="AJR21487.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(8181..8975)
                     /gene="aphA-3"
                     /label=aphA-3
     regulatory      complement(9178..9392)
                     /regulatory_class="terminator"
                     /note="CaMV 35S terminator"
     misc_feature    9400..9424
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    9502..9676
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9716..10741)
                     /codon_start=1
                     /gene="hpt"
                     /product="hygromycin phosphotransferase"
                     /function="hygromycin resistance"
                     /label=hpt
                     /protein_id="AJR21488.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(9716..10741)
                     /gene="hpt"
                     /label=hpt
     regulatory      complement(10765..11534)
                     /regulatory_class="promoter"
                     /gene="CaMV 35S"
     gene            complement(10765..11534)
                     /gene="CaMV 35S"
                     /label=CaMV 35S
     promoter        complement(10809..11486)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    11677..11698
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11713..11743
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11751..11767
                     /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     11775..11791
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"