LOCUS       Exported                7240 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCLEAN-G184, complete sequence.
ACCESSION   JN207920
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7240)
  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 7240)
  AUTHORS   Thole V, Vain P.
  TITLE     Direct Submission
  JOURNAL   Submitted (04-JUL-2011) Department of Crop Genetics, John Innes 
            Centre, Norwich Research Park, Norwich, Norfolk NR4 7UH, United 
            Kingdom
REFERENCE   3  (bases 1 to 7240)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7240
                     /organism="Binary vector pCLEAN-G184"
                     /mol_type="other DNA"
                     /db_xref="taxon:1116802"
     misc_feature    1..4224
                     /note="derived from binary vector pCLEAN-G182 in INSD 
                     accession EU186092"
     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="AET86651.1"
                     /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(2536..4098)
                     /note="CaMV35S:mgfp5-ER:SPA expression unit inserted in 
                     vector backbone outside the left border sequence of pCLEAN 
                     T-DNA region"
     regulatory      complement(2536..2832)
                     /regulatory_class="terminator"
                     /note="SPA-ter; soybean-derived poly(A) signal"
     CDS             complement(2853..3644)
                     /codon_start=1
                     /gene="mgfp5-ER"
                     /product="mGFP5-ER"
                     /label=mgfp5-ER
                     /note="modified H46Y green fluorescent protein mgfp5-ER"
                     /protein_id="AET86652.1"
                     /translation="MKTNLFLFLIFSLLLSLSSAEFSKGEELFTGVVPILVELDGDVNG
                     YKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDF
                     FKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLE
                     YNYNSHNVYIMADKQKNGIKANFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYL
                     STQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKHDEL"
     gene            complement(2853..3644)
                     /gene="mgfp5-ER"
                     /label=mgfp5-ER
     CDS             complement(2868..3578)
                     /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(3671..4098)
                     /regulatory_class="promoter"
                     /note="35S promoter from CaMV"
     promoter        complement(3677..4021)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    4137..4161
                     /note="nopaline-type T-DNA left border sequence No.1 
                     belonging to a double left border sequence separated by a 
                     spacer sequence from left border No.2"
     misc_feature    4162..4187
                     /note="spacer sequence separating left border No.1 and No.2
                     of the double left border sequence"
     misc_feature    4188..4212
                     /note="nopaline-type T-DNA left border No.2 belonging to a 
                     double left border sequence separated by a spacer sequence 
                     from left border No.1"
     misc_feature    4225..7113
                     /label=derived from binary vector pGVT5
                     /note="derived from binary vector pGVT5"
     regulatory      complement(4256..4548)
                     /regulatory_class="terminator"
                     /note="SPA-ter; soybean-derived poly(A) signal"
     CDS             complement(join(4623..6049,6239..6623))
                     /codon_start=1
                     /gene="gusA"
                     /product="beta-glucuronidase"
                     /label=gusA
                     /protein_id="AET86653.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIGQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            complement(4623..6623)
                     /gene="gusA"
                     /label=gusA
     intron          complement(6050..6238)
                     /gene="gusA"
                     /note="modified 2nd intron (IV2) of ST-LS1 gene"
     regulatory      complement(6655..7082)
                     /regulatory_class="promoter"
                     /note="35S promoter from CaMV"
     promoter        complement(6661..7005)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    7158..7182
                     /label=nopaline-type T-DNA right border sequence
                     /note="nopaline-type T-DNA right border sequence"
     misc_feature    7183..7206
                     /label=right border overdrive sequence
                     /note="right border overdrive sequence"