LOCUS       Exported               11846 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCAMBIA-1305.1, complete sequence.
ACCESSION   AF354045
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11846)
  AUTHORS   Jefferson RA, Kilian A, Keese PK.
  TITLE     Microbial genes for secreted beta-glucuronidases, gene products and 
            uses thereof
  JOURNAL   Patent: PCT (PCT/US98/19217)-A 09-SEP-1998; CAMBIA; Molecular 
            Technologies; Canberra; Australia;
REFERENCE   2  (bases 1 to 11846)
  AUTHORS   Nguyen TA, Jefferson RA.
  TITLE     Binary vector pCAMBIA-1305.1
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 11846)
  AUTHORS   Nguyen TA, Jefferson RA.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-FEB-2001) Molecular Technologies, CAMBIA, GPO Box 
            3200, Canberra, ACT 2601, Australia
REFERENCE   4  (bases 1 to 11846)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11846
                     /organism="Binary vector pCAMBIA-1305.1"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:156490"
     CDS             join(2..16,207..2054)
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=beta-glucuronidase
                     /note="GUSPlus-His6"
                     /protein_id="AAK29426.1"
                     /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="catalase intron"
     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="polyA_signal_sequence"
                     /note="poly(A) signal from nopaline synthase"
     misc_feature    2354..2378
                     /label=T-DNA right border
                     /note="T-DNA right border"
     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=pVS1 rep
                     /note="pVS1 rep"
     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
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /note="kanamycin resistance gene"
                     /protein_id="AAK29424.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    8632..8657
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    8632..8656
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    8734..8908
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     polyA_site      8798..8805
     CDS             complement(8948..9973)
                     /codon_start=1
                     /product="hygromycin phosphotransferase"
                     /label=hygromycin phosphotransferase
                     /note="hygromycin resistance gene"
                     /protein_id="AAK29425.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     regulatory      complement(10009..10816)
                     /regulatory_class="promoter"
                     /note="CaMV35Sx2; CaMV35S eukaryotic promoter 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      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"
     CDS             11019..11252
                     /codon_start=1
                     /product="lacZ alpha fragment"
                     /label=lacZ alpha fragment
                     /note="beta-galactosidase alpha fragment"
                     /protein_id="AAK29423.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     misc_feature    11033..11089
                     /label=pUC18 polylinker multiple cloning site
                     /note="pUC18 polylinker multiple cloning site"
     primer_bind     complement(11093..11109)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      11303..11840
                     /regulatory_class="promoter"
                     /note="CaMV35S eukaryotic promoter"
     promoter        11486..11831
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"