LOCUS       Exported                8663 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pGPro5, complete sequence.
ACCESSION   JN593326
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8663)
  AUTHORS   Thilmony R.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-AUG-2011) Crop Improvement and Utilization research 
            Unit, USDA-ARS-WRRC, 800 Buchanan Street, Albany, CA 94710-1105, USA
REFERENCE   2  (bases 1 to 8663)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8663
                     /organism="Binary vector pGPro5"
                     /mol_type="other DNA"
                     /db_xref="taxon:1126005"
     misc_feature    1..25
                     /label=right border sequence for T-DNA
                     /note="right border sequence for T-DNA"
     protein_bind    264..285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        300..330
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    338..354
                     /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     362..378
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        399..417
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    430..537
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     454..470
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(504..520)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             join(540..629,820..2667)
                     /codon_start=1
                     /gene="GRP-GUSPlus-His6"
                     /product="GRP-GUSPlus-His6"
                     /label=GRP-GUSPlus-His6
                     /note="codon-optimized beta-glucuronidase from 
                     Staphylococcus sp. with rice glycine rich protein (GRP) 
                     signal peptide to target the protein for extracellular 
                     secretion; C-terminal HexaHis tag"
                     /protein_id="AEW22790.1"
                     /translation="MATTKHLALAILVLLSIGMTTSARTLLDLRNRRTSLYPINTETRG
                     VFDLNGVWNFKLDYGKGLEEKWYESKLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYER
                     EFTVPAYLKDQRIVLRFGSATHKAIVYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVT
                     VAVDNILDDSTLPVGLYSERHEEGLGKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVED
                     ISVVTDFNGPTGTVTYTVDFQGKAETVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEP
                     LNTYLYQIKVELVNDGLTIDVYEEPFGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPIN
                     GRGFNEASNVMDFNILKWIGANSFRTAHYPYSEELMRLADREGLVVIDETPAVGVHLNF
                     MATTGLGEGSERVSTWEKIRTFEHHQDVLRELVSRDKNHPSVVMWSIANEAATEEEGAY
                     EYFKPLVELTKELDPQKRPVTIVLFVMATPETDKVAELIDVIALNRYNGWYFDGGDLEA
                     AKVHLRQEFHAWNKRCPGKPIMITEYGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFD
                     EFENFVGEQAWNFADFATSQGVMRVQGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYK
                     NASHHHHHHV"
     gene            540..2667
                     /gene="GRP-GUSPlus-His6"
                     /label=GRP-GUSPlus-His6
     CDS             540..620
                     /codon_start=1
                     /product="signal sequence from Oryza sativa glycine-rich 
                     cell wall protein"
                     /label=GRP signal sequence
                     /translation="MATTKHLALAILVLLSIGMTTSARTLL"
     CDS             join(621..629,820..2637)
                     /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          630..819
                     /gene="GRP-GUSPlus-His6"
                     /note="catalase intron"
     CDS             2644..2661
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2693..2945
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      2718..2948
                     /regulatory_class="terminator"
                     /note="nos transcription terminator"
     misc_feature    2955..2988
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    2955..2988
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      2995..4713
                     /regulatory_class="promoter"
                     /note="Oryza sativa japonica group cv. Nipponbare ubiquitin
                     2 promoter"
     promoter        2995..4704
                     /gene="RUBQ2"
                     /label=Ubi promoter
                     /note="promoter for the rice (Oryza sativa) polyubiquitin 
                     gene RUBQ2"
     intron          3743..4704
                     /note="Oryza sativa japonica group cv. Nipponbare ubiquitin
                     2 intron"
     CDS             4725..5750
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin phosphotransferase"
                     /label=hptII
                     /note="hygromycin resistance marker"
                     /protein_id="AEW22791.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            4725..5750
                     /gene="hptII"
                     /label=hptII
     misc_feature    5782..5815
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    5782..5815
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     polyA_signal    5830..6004
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    6081..6106
                     /label=left border sequence for T-DNA
                     /note="left border sequence for T-DNA"
     misc_feature    6082..6106
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    6114..6138
                     /label=left border sequence for T-DNA
                     /note="left border sequence for T-DNA"
     misc_feature    6116..6138
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     misc_feature    6139..8663
                     /label=pGreenII 0000 vector sequence
                     /note="pGreenII 0000 vector sequence"
     rep_origin      6273..6708
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6999..7814
                     /codon_start=1
                     /gene="aph(3)-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANVVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      7985..8573
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"