LOCUS       Exported                8588 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pGPro8, complete sequence.
ACCESSION   JN593327
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8588)
  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 8588)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8588
                     /organism="Binary vector pGPro8"
                     /mol_type="other DNA"
                     /db_xref="taxon:1126006"
     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..554,745..2592)
                     /codon_start=1
                     /gene="GUSPlus-His6"
                     /product="GUSPlus-His6"
                     /label=GUSPlus-His6
                     /note="codon-optimized beta-glucuronidase from 
                     Staphylococcus sp.; C-terminal HexaHis tag"
                     /protein_id="AEW22792.1"
                     /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES
                     KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI
                     VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL
                     GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE
                     TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP
                     FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR
                     TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH
                     QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF
                     VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE
                     YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV
                     QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKNASHHHHHHV"
     gene            540..2592
                     /gene="GUSPlus-His6"
                     /label=GUSPlus-His6
     CDS             join(540..554,745..2562)
                     /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          555..744
                     /gene="GUSPlus-His6"
                     /note="catalase intron"
     CDS             2569..2586
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2618..2870
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      2643..2873
                     /regulatory_class="terminator"
                     /note="nos transcription terminator"
     misc_feature    2880..2913
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    2880..2913
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      2920..4638
                     /regulatory_class="promoter"
                     /note="Oryza sativa japonica group cv. Nipponbare ubiquitin
                     2 promoter"
     promoter        2920..4629
                     /gene="RUBQ2"
                     /label=Ubi promoter
                     /note="promoter for the rice (Oryza sativa) polyubiquitin 
                     gene RUBQ2"
     intron          3668..4629
                     /note="Oryza sativa japonica group cv. Nipponbare ubiquitin
                     2 intron"
     CDS             4650..5675
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin phosphotransferase"
                     /label=hptII
                     /note="hygromycin resistance marker"
                     /protein_id="AEW22793.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            4650..5675
                     /gene="hptII"
                     /label=hptII
     misc_feature    5707..5740
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    5707..5740
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      5741..5956
                     /regulatory_class="terminator"
                     /note="CaMV 35S transcription terminator"
     polyA_signal    5755..5929
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    6006..6031
                     /label=left border sequence for T-DNA
                     /note="left border sequence for T-DNA"
     misc_feature    6007..6031
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    6039..6063
                     /label=left border sequence for T-DNA
                     /note="left border sequence for T-DNA"
     misc_feature    6041..6063
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     misc_feature    6064..8588
                     /label=pGreenII 0000 vector sequence
                     /note="pGreenII 0000 vector sequence"
     rep_origin      6198..6633
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6924..7739
                     /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      7910..8498
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"