LOCUS       Exported                8809 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pGPro2, complete sequence.
ACCESSION   EU147786
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8809)
  AUTHORS   Thilmony R, Guttman M, Chiniquy D, Blechl A.
  TITLE     pGPro1, A Novel Binary Vector for Monocot Promoter Characterization
  JOURNAL   Plant Mol. Biol. Rep. 24, 57-69 (2006)
REFERENCE   2  (bases 1 to 8809)
  AUTHORS   Thilmony R, Guttman M, Chiniquy D, Blechl A.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-SEP-2007) Crop Improvement and Utilization Research 
            Unit, USDA-ARS Western Regional Research Center, 800 Buchanan 
            Street, Albany, CA 94710-1105, USA
REFERENCE   3  (bases 1 to 8809)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8809
                     /organism="Binary vector pGPro2"
                     /mol_type="other DNA"
                     /db_xref="taxon:473624"
     CDS             9..2576
                     /codon_start=1
                     /gene="gusA-eGFP"
                     /product="gusA-eGFP"
                     /label=gusA-eGFP
                     /note="beta-glucuronidase-enhanced green fluorescent fusion
                     protein"
                     /protein_id="ABV71994.1"
                     /translation="MVDLTSLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQ
                     ESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWV
                     NNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQ
                     SYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELR
                     DADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAV
                     KGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAE
                     EMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKE
                     LIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHT
                     DTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAG
                     LHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIF
                     TRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQASGAGGAGMVSKGEELFTGVVPILVEL
                     DGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDH
                     MKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNI
                     LGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLL
                     PDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     gene            9..2576
                     /gene="gusA-eGFP"
                     /label=gusA-eGFP
     CDS             27..1832
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="LRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAIA
                     VPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEVM
                     EHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHDF
                     FNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQV
                     VATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQFL
                     INHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDWA
                     DEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARDK
                     NHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISDL
                     FDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYT
                     DMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKP
                     KSAAFLLQKRWTGMNFGEKPQQGGKQ"
     CDS             1857..2576
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     regulatory      2583..2887
                     /regulatory_class="terminator"
                     /note="nos transcription terminator"
     CDS             2583..2600
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2632..2884
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2894..4324
                     /label=loxP attachment site for the Cre recombinase
                     /note="loxP attachment site for the Cre recombinase"
     protein_bind    2894..2927
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      2934..4324
                     /regulatory_class="promoter"
                     /note="Oryza sativa (japonica cultivar-group) cv. 
                     Nipponbare actin promoter derived from Os03g50890"
     CDS             3775..3792
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4340..5365
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin phosphotransferase"
                     /label=hptII
                     /note="plant selectable marker"
                     /protein_id="ABV71995.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            4340..5365
                     /gene="hptII"
                     /label=hptII
     misc_feature    5397..5430
                     /label=loxP attachment site for the Cre recombinase
                     /note="loxP attachment site for the Cre recombinase"
     protein_bind    5397..5430
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      5445..5690
                     /regulatory_class="terminator"
                     /note="CaMV 35S transcription terminator"
     polyA_signal    5445..5619
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5696..5721
                     /label=left border sequence for T-DNA (second copy)
                     /note="left border sequence for T-DNA (second copy)"
     misc_feature    5697..5721
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    5724..8809
                     /label=pGreenII 0000 vector sequence
                     /note="pGreenII 0000 vector sequence"
     misc_feature    5731..5753
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      5888..6323
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6614..7429
                     /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      7600..8188
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    8279..8303
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    8542..8563
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8578..8608
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8616..8632
                     /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     8640..8656
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        8677..8695
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_feature    8708..6
                     /label=MCS
                     /note="pBluescript multiple cloning site"
     primer_bind     8732..8748
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8782..8798)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"