LOCUS       Exported                7171 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCLEAN-G156, complete sequence.
ACCESSION   JN654719
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7171)
  AUTHORS   Thole V, Worland B, Snape JW, Vain P.
  TITLE     The pCLEAN dual binary vector system for Agrobacterium-mediated 
            plant transformation
  JOURNAL   Plant Physiol. 145 (4), 1211-1219 (2007)
  PUBMED    17932303
REFERENCE   2  (bases 1 to 7171)
  AUTHORS   Thole V, Vain P.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-SEP-2011) Department of Crop Genetics, John Innes 
            Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH, United 
            Kingdom
REFERENCE   3  (bases 1 to 7171)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7171
                     /organism="Binary vector pCLEAN-G156"
                     /mol_type="other DNA"
                     /db_xref="taxon:1136950"
     misc_feature    1..4177
                     /note="derived from binary vector pCLEAN-G130 in INSD 
                     accession number EU186091"
     rep_origin      1..720
                     /label=pUC-ori
                     /note="pUC-ori"
     rep_origin      complement(61..649)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(820..1635)
                     /codon_start=1
                     /gene="nptI"
                     /product="kanamycin resistance protein"
                     /label=nptI
                     /protein_id="AEY82453.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(820..1635)
                     /gene="nptI"
                     /label=nptI
     rep_origin      1921..2473
                     /label=pSa-ori
                     /note="pSa-ori"
     rep_origin      1926..2361
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    complement(2538..4100)
                     /note="CaMV:mgfp5-ER:SPA expression unit inserted in vector
                     backbone outside the left border sequence of pCLEAN T-DNA 
                     region"
     regulatory      complement(2538..2834)
                     /regulatory_class="terminator"
                     /note="SPA-ter; soybean-derived poly(A) signal"
     CDS             complement(2855..3646)
                     /codon_start=1
                     /gene="mgfp5-ER"
                     /product="mGFP5-ER"
                     /label=mgfp5-ER
                     /note="modified green fluorescent protein mgfp5-ER in INSD 
                     accession number U87974 containing one nucleotide variation
                     resulting in an H46Y amino acid change"
                     /protein_id="AEY82455.1"
                     /translation="MKTNLFLFLIFSLLLSLSSAEFSKGEELFTGVVPILVELDGDVNG
                     YKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDF
                     FKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLE
                     YNYNSHNVYIMADKQKNGIKANFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYL
                     STQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKHDEL"
     gene            complement(2855..3646)
                     /gene="mgfp5-ER"
                     /label=mgfp5-ER
     CDS             complement(2870..3580)
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="SKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKAN
                     FKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     regulatory      complement(3673..4100)
                     /regulatory_class="promoter"
                     /note="35S promoter of CaMV"
     promoter        complement(3679..4023)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    4141..4165
                     /label=nopaline-type T-DNA left border sequence
                     /note="nopaline-type T-DNA left border sequence"
     misc_feature    4178..7066
                     /label=derived from binary vector pGVT5
                     /note="derived from binary vector pGVT5"
     regulatory      complement(4209..4501)
                     /regulatory_class="terminator"
                     /note="SPA-ter; soybean-derived poly(A) signal"
     CDS             complement(join(4576..6002,6192..6576))
                     /codon_start=1
                     /gene="gusA"
                     /product="beta-glucuronidase"
                     /label=gusA
                     /protein_id="AEY82454.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIGQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            complement(4576..6576)
                     /gene="gusA"
                     /label=gusA
     intron          complement(6003..6191)
                     /gene="gusA"
                     /note="modified 2nd intron (IV2) of ST-LS1 gene"
     regulatory      complement(6608..7035)
                     /regulatory_class="promoter"
                     /note="35S promoter of CaMV"
     promoter        complement(6614..6958)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    7111..7135
                     /label=nopaline-type T-DNA right border sequence
                     /note="nopaline-type T-DNA right border sequence"
     misc_feature    7136..7159
                     /label=right border overdrive sequence
                     /note="right border overdrive sequence"