LOCUS       Exported               11951 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pJZ253, complete sequence.
ACCESSION   KP244679
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11951)
  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 11951)
  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 11951)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11951
                     /organism="Binary vector pJZ253"
                     /mol_type="other DNA"
                     /db_xref="taxon:1622250"
     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    1436..1732
                     /label=similar to Leptinotarsa decemlineata actin gene
                     /note="similar to Leptinotarsa decemlineata actin gene"
     intron          1740..1940
                     /gene="St-GA20 ox"
                     /number=1
                     /note="first intron from potato GA20 oxidase gene"
     gene            1740..1940
                     /gene="St-GA20 ox"
                     /label=St-GA20 ox
                     /note="GA20 oxidase"
     misc_feature    complement(1948..2244)
                     /label=similar to Leptinotarsa decemlineata actin gene
                     /note="similar to Leptinotarsa decemlineata actin gene"
     terminator      2254..2961
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     regulatory      2254..2960
                     /regulatory_class="terminator"
                     /note="octopine synthase gene terminator"
     promoter        complement(3002..3020)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3053..3069)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   3271..3294
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    3272..3296
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      4356..5356
                     /label=pVS1
                     /note="pVS1"
     CDS             4596..5225
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5659..6726
                     /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      6792..6986
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    7330..7470
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(7656..8244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      7759..8040
                     /label=pBR325
                     /note="pBR325"
     CDS             complement(8331..9125)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /function="kanamycin resistance"
                     /label=aphA-3
                     /protein_id="AJR21485.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(8331..9125)
                     /gene="aphA-3"
                     /label=aphA-3
     regulatory      complement(9328..9542)
                     /regulatory_class="terminator"
                     /note="CaMV 35S terminator"
     misc_feature    9550..9574
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    9652..9826
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9866..10891)
                     /codon_start=1
                     /gene="hpt"
                     /product="hygromycin phosphotransferase"
                     /function="hygromycin resistance"
                     /label=hpt
                     /protein_id="AJR21486.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(9866..10891)
                     /gene="hpt"
                     /label=hpt
     regulatory      10915..11684
                     /regulatory_class="promoter"
                     /gene="CaMV 35S"
     gene            10915..11684
                     /gene="CaMV 35S"
                     /label=CaMV 35S
     promoter        complement(10959..11636)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    11827..11848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11863..11893
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11901..11917
                     /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     11925..11941
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"