LOCUS       Exported               13701 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pGFPGUSplus, complete sequence.
ACCESSION   EF546437
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13701)
  AUTHORS   Vickers CE, Schenk PM, Li D, Mullineaux PM, Gresshoff PM.
  TITLE     pGFPGUSPlus, a new binary vector for gene expression studies and 
            optimising transformation systems in plants
  JOURNAL   Biotechnol. Lett. 29 (11), 1793-1796 (2007)
  PUBMED    17687623
REFERENCE   2  (bases 1 to 13701)
  AUTHORS   Vickers CE, Schenk PM, Mullineaux PM, Gresshoff PM.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-APR-2007) Biological Sciences, The University of 
            Essex, Wivenhoe Park, Colchester, Essex CO43SQ, England
REFERENCE   3  (bases 1 to 13701)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13701
                     /organism="Binary vector pGFPGUSplus"
                     /mol_type="other DNA"
                     /db_xref="taxon:436309"
     CDS             join(2..16,207..2054)
                     /codon_start=1
                     /product="beta-glucuronidase GUSPlus-His6"
                     /label=beta-glucuronidase GUSPlus-His6
                     /note="similar to GenBank accession number AAK29426"
                     /protein_id="ABQ22951.1"
                     /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES
                     KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI
                     VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL
                     GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE
                     TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP
                     FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR
                     TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH
                     QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF
                     VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE
                     YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV
                     QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKNASHHHHHHV"
     CDS             join(2..16,207..2024)
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=GUSPlus(TM)
                     /note="codon-optimized Staphylococcus gusA gene with a 
                     catalase intron to ensure expression in plants but not 
                     bacteria"
                     /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES
                     KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI
                     VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL
                     GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE
                     TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP
                     FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR
                     TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH
                     QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF
                     VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE
                     YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV
                     QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN"
     intron          17..206
                     /note="from catalase"
     CDS             2031..2048
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2080..2332
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      2083..2335
                     /regulatory_class="terminator"
                     /note="from nopaline synthase (NOS)"
     regulatory      2099..2104
                     /regulatory_class="polyA_signal_sequence"
                     /note="from nopaline synthase"
     misc_feature    2338..2487
                     /label=T-DNA right border (RB)
                     /note="T-DNA right border (RB)"
     repeat_region   2354..2378
                     /label=right border repeat (RBR)
                     /note="right border repeat (RBR)"
     misc_feature    2354..2378
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    3439..4439
                     /label=sta region from pVS1 plasmid
                     /note="sta region from pVS1 plasmid"
     CDS             3678..4307
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4741..5808
                     /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      5032..6032
                     /label=from pVS1
                     /note="from pVS1"
     rep_origin      5874..6068
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6412..6552
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    6442..6702
                     /label=pBR322 bom site
                     /note="pBR322 bom site"
     rep_origin      complement(6738..7326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6842..7122
                     /label=pBR322 ori
                     /note="pBR322 ori"
     CDS             complement(7413..8207)
                     /codon_start=1
                     /gene="Aph"
                     /product="neomycin phosphotransferase"
                     /label=Aph
                     /note="kanamycin resistance protein from pIG121Hm in 
                     GenBank accession number AAK29424.1"
                     /protein_id="ABQ22952.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(7413..8207)
                     /gene="Aph"
                     /label=Aph
     misc_feature    8632..8657
                     /label=T-DNA left border (LB)
                     /note="T-DNA left border (LB)"
     misc_feature    8632..8656
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      complement(8660..8922)
                     /regulatory_class="terminator"
                     /note="CaMV 35S 3 terminator region"
     polyA_signal    8734..8908
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     regulatory      complement(8798..8805)
                     /regulatory_class="polyA_signal_sequence"
     CDS             complement(8948..9973)
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin phosphotransferase"
                     /function="hygromycin resistance"
                     /label=hptII
                     /note="APH in GenBank accession number AAK29425.1"
                     /protein_id="ABQ22953.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(8948..9973)
                     /gene="hptII"
                     /label=hptII
     regulatory      complement(10009..10816)
                     /regulatory_class="promoter"
                     /note="2xCaMV35S; from CaMV35S, with duplicated enhancer 
                     region"
     promoter        complement(10041..10718)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    10909..10930
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      complement(10938..11018)
                     /regulatory_class="promoter"
                     /note="PlacZ prokaryotic promoter"
     promoter        10945..10975
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10983..10999
                     /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     11007..11023
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(11041..11293)
                     /regulatory_class="terminator"
                     /note="from nopaline synthase (NOS)"
     terminator      11041..11293
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(11339..12058)
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=green fluorescent protein
                     /note="includes S65T mutation"
                     /protein_id="ABQ22954.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      complement(12086..12920)
                     /regulatory_class="promoter"
                     /note="2xCaMV35S; from CaMV35S, with duplicated enhancer 
                     region"
     promoter        complement(12088..12433)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(12948..12964)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      13267..13688
                     /regulatory_class="promoter"
                     /note="from CaMV35S"
     promoter        13341..13686
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"