LOCUS       Exported               11921 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCAMBIA-1305.2, complete sequence.
ACCESSION   AF354046
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11921)
  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 11921)
  AUTHORS   Nguyen TA, Jefferson RA.
  TITLE     Binary pCAMBIA-1305.2
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 11921)
  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 11921)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11921
                     /organism="Binary vector pCAMBIA-1305.2"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:156493"
     CDS             join(2..91,282..2129)
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=beta-glucuronidase
                     /note="GRP-GusPlus-His6; GRP signal peptide from 2-81 
                     (glycine-rich protein) for secretion of GUSPlus-His6"
                     /protein_id="AAK29427.1"
                     /translation="MATTKHLALAILVLLSIGMTTSARTLLDLRNRRTSLYPINTETRG
                     VFDLNGVWNFKLDYGKGLEEKWYESKLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYER
                     EFTVPAYLKDQRIVLRFGSATHKAIVYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVT
                     VAVDNILDDSTLPVGLYSERHEEGLGKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVED
                     ISVVTDFNGPTGTVTYTVDFQGKAETVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEP
                     LNTYLYQIKVELVNDGLTIDVYEEPFGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPIN
                     GRGFNEASNVMDFNILKWIGANSFRTAHYPYSEELMRLADREGLVVIDETPAVGVHLNF
                     MATTGLGEGSERVSTWEKIRTFEHHQDVLRELVSRDKNHPSVVMWSIANEAATEEEGAY
                     EYFKPLVELTKELDPQKRPVTIVLFVMATPETDKVAELIDVIALNRYNGWYFDGGDLEA
                     AKVHLRQEFHAWNKRCPGKPIMITEYGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFD
                     EFENFVGEQAWNFADFATSQGVMRVQGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYK
                     NASHHHHHHV"
     CDS             2..82
                     /codon_start=1
                     /product="signal sequence from Oryza sativa glycine-rich 
                     cell wall protein"
                     /label=GRP signal sequence
                     /translation="MATTKHLALAILVLLSIGMTTSARTLL"
     CDS             join(83..91,282..2099)
                     /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="DLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYESKL
                     TDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAIVY
                     VNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGLGK
                     VIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAETV
                     KVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEPFG
                     VRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFRTA
                     HYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHHQD
                     VLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLFVM
                     ATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITEYG
                     ADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRVQG
                     NKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN"
     intron          92..281
                     /note="catalase intron"
     CDS             2106..2123
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2155..2407
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      2158..2410
                     /regulatory_class="polyA_signal_sequence"
                     /note="poly(A) signal from nopaline synthase"
     misc_feature    2429..2453
                     /label=T-DNA right border
                     /note="T-DNA right border"
     misc_feature    3514..4514
                     /label=sta region from pVS1 plasmid
                     /note="sta region from pVS1 plasmid"
     CDS             3753..4382
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4816..5883
                     /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      5107..6107
                     /label=pVS1 rep
                     /note="pVS1 rep"
     rep_origin      5949..6143
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6487..6627
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    6517..6777
                     /label=pBR322 bom site
                     /note="pBR322 bom site"
     rep_origin      complement(6813..7401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      6917..7197
                     /label=pBR322 ori
                     /note="pBR322 ori"
     CDS             complement(7488..8282)
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /note="kanamycin resistance gene; npt II gene amplified 
                     from pIG121Hm; NPT II"
                     /protein_id="AAK29428.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    8707..8732
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    8707..8731
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    8809..8983
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     polyA_site      8873..8880
     CDS             complement(9023..10048)
                     /codon_start=1
                     /product="hygromycin phosphotransferase"
                     /label=hygromycin phosphotransferase
                     /note="HPT II"
                     /protein_id="AAK29429.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(10116..10793)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     regulatory      complement(10891..11084)
                     /regulatory_class="promoter"
                     /note="CaMV35Sx2; CaMV35S eukaryotic promoter with 
                     duplicated enhancer region"
     protein_bind    10984..11005
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      11013..11090
                     /regulatory_class="promoter"
                     /note="PlacZ prokaryotic promoter"
     promoter        11020..11050
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11058..11074
                     /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     11082..11098
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             11094..11327
                     /codon_start=1
                     /product="beta-galactosidase alpha fragment"
                     /label=beta-galactosidase alpha fragment
                     /note="lacZ alpha"
                     /protein_id="AAK29430.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     misc_feature    11108..11164
                     /label=pUC18 polylinker multiple cloning site
                     /note="pUC18 polylinker multiple cloning site"
     primer_bind     complement(11168..11184)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      11378..11915
                     /regulatory_class="promoter"
                     /note="CaMV35S eukaryotic promoter"
     promoter        11561..11906
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"