LOCUS       Exported                7698 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pGPro4, complete sequence.
ACCESSION   JN593324
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7698)
  AUTHORS   Chen GQ, Thilmony R, Lin J-T.
  TITLE     Transformation of Lesquerella fendleri with the new binary vector 
            pGPro4-35S
  JOURNAL   Online J Biol Sci 11 (3), 90-95 (2012)
REFERENCE   2  (bases 1 to 7698)
  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   3  (bases 1 to 7698)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7698
                     /organism="Binary vector pGPro4"
                     /mol_type="other DNA"
                     /db_xref="taxon:1126003"
     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             540..3107
                     /codon_start=1
                     /gene="gusA-eGFP"
                     /product="beta-glucuronidase-enhanced green fluorescent 
                     fusion protein"
                     /label=gusA-eGFP
                     /note="bifunctional reporter"
                     /protein_id="AEW22786.1"
                     /translation="MVDLTSLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQ
                     ESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWV
                     NNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQ
                     SYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELR
                     DADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAV
                     KGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAE
                     EMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKE
                     LIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHT
                     DTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAG
                     LHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIF
                     TRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQASGAGGAGMVSKGEELFTGVVPILVEL
                     DGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDH
                     MKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNI
                     LGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLL
                     PDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     gene            540..3107
                     /gene="gusA-eGFP"
                     /label=gusA-eGFP
     CDS             558..2363
                     /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             2388..3107
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             3114..3131
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      3163..3415
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      3188..3418
                     /regulatory_class="terminator"
                     /note="nos transcription terminator"
     misc_feature    3425..3458
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    3425..3458
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      3473..3745
                     /regulatory_class="promoter"
                     /note="Agrobacterium nopaline synthase (nos) promoter"
     promoter        3545..3724
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             3760..4785
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin phosphotransferase"
                     /label=hptII
                     /note="hygromycin resistance marker"
                     /protein_id="AEW22787.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            3760..4785
                     /gene="hptII"
                     /label=hptII
     misc_feature    4817..4850
                     /label=loxP attachment site for Cre recombinase
                     /note="loxP attachment site for Cre recombinase"
     protein_bind    4817..4850
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      4851..5066
                     /regulatory_class="terminator"
                     /note="CaMV 35S transcription terminator"
     polyA_signal    4865..5039
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    5116..5141
                     /label=left border sequence for T-DNA
                     /note="left border sequence for T-DNA"
     misc_feature    5117..5141
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    5149..5173
                     /label=left border sequence for T-DNA
                     /note="left border sequence for T-DNA"
     misc_feature    5151..5173
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     misc_feature    5174..7698
                     /label=pGreenII 0000 vector sequence
                     /note="pGreenII 0000 vector sequence"
     rep_origin      5308..5743
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     CDS             6034..6849
                     /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      7020..7608
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"