LOCUS       Exported                9737 bp ds-DNA    circular SYN 11-1-2016
DEFINITION  Agrobacterium binary vector for plant transformation, with 
            kanamycin-resistance and secreted GUSPlus(TM) genes.
ACCESSION   .
VERSION     .
KEYWORDS    pCAMBIA0305.2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9737)
  AUTHORS   Cambia
  TITLE     Direct Submission
COMMENT     The GenBank record was corrected by inserting a G at position 4815.
FEATURES             Location/Qualifiers
     source          1..9737
                     /organism="synthetic DNA construct"
                     /lab_host="Plant Cells"
                     /mol_type="other DNA"
     CDS             2..82
                     /codon_start=1
                     /product="signal sequence from Oryza sativa glycine-rich 
                     cell wall protein"
                     /note="GRP signal sequence"
                     /translation="MATTKHLALAILVLLSIGMTTSARTLL"
     CDS             join(83..91,282..2099)
                     /codon_start=1
                     /gene="
                     "
                     /product="beta-glucuronidase"
                     /note="GUSPlus(TM)"
                     /note="codon-optimized Staphylococcus gusA gene with a 
                     catalase intron to ensure expression in plants but not 
                     bacteria"
                     /protein_id="
                     "
                     /translation="DLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYESKL
                     TDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAIVY
                     VNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGLGK
                     VIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAETV
                     KVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEPFG
                     VRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFRTA
                     HYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHHQD
                     VLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLFVM
                     ATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITEYG
                     ADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRVQG
                     NKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN"
     intron          92..281
                     /note="cat1 intron"
                     /note="castor bean catalase intron, modified"
     CDS             2106..2123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     terminator      2155..2407
                     /note="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2429..2453
                     /note="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3753..4382
                     /codon_start=1
                     /product="stability protein from Pseudomonas plasmid pVS1"
                     /note="pVS1 StaA"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4811..5884
                     /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      5950..6144
                     /note="pVS1 oriV"
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6488..6628
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6814..7402)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7489..8283)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /protein_id="
                     "
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    8708..8732
                     /note="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        8836..8866
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    8874..8890
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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     8898..8914
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             8910..9143
                     /codon_start=1
                     /gene="lacZ (truncated)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /protein_id="
                     "
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     misc_feature    8924..8980
                     /gene="
                     "
                     /note="MCS"
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(8984..9000)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9377..9722
                     /note="CaMV 35S promoter"
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"