LOCUS       Exported               11406 bp ds-DNA    circular SYN 11-1-2016
DEFINITION  Agrobacterium binary vector for expressing dsRNA, with two multiple 
            cloning sites separated by an intron.
ACCESSION   AY310901
VERSION     .
KEYWORDS    pFGC5941
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11406)
  AUTHORS   Kerschen A, Napoli CA, Jorgensen RA, Muller AE.
  TITLE     Effectiveness of RNA interference in transgenic plants.
  JOURNAL   FEBS Lett. 2004;566:223-8.
  PUBMED    15147899
REFERENCE   2  (bases 1 to 11406)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     The GenBank sequence was corrected by inserting a G at position 
            7514.
FEATURES             Location/Qualifiers
     source          1..11406
                     /organism="synthetic DNA construct"
                     /lab_host="Plant Cells"
                     /mol_type="other DNA"
     misc_feature    1..25
                     /note="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      111..363
                     /note="MAS terminator"
                     /note="mannopine synthase terminator"
     CDS             complement(373..924)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /function="
                     "
                     /note="BlpR"
                     /note="confers resistance to bialophos or phosphinothricin"
                     /protein_id="
                     "
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(930..1310)
                     /note="MAS promoter"
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     promoter        2305..2650
                     /note="CaMV 35S promoter"
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    2680..2734
                     /note="TMV Omega"
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     misc_feature    2734..2764
                     /note="MCS 1"
                     /note="multiple cloning site 1"
     intron          2767..4115
                     /gene="chsA"
                     /number="
                     "
                     /note="chsA intron"
                     /note="chalcone synthase A intron from Petunia hybrida"
     misc_feature    4122..4160
                     /note="MCS 2"
                     /note="multiple cloning site"
     terminator      4177..4884
                     /note="OCS terminator"
                     /note="octopine synthase terminator"
     misc_feature    5128..5152
                     /note="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             6452..7081
                     /codon_start=1
                     /product="stability protein from Pseudomonas plasmid pVS1"
                     /note="pVS1 StaA"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7510..8583
                     /codon_start=1
                     /product="replication protein from Pseudomonas plasmid 
                     pVS1"
                     /note="pVS1 RepA"
                     /note="
                     "
                     /protein_id="
                     "
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      8649..8843
                     /note="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    9187..9327
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9513..10101)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10188..10982)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /protein_id="
                     "
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"