LOCUS       Exported                8114 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCLEAN-G265, complete sequence.
ACCESSION   KF974574
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8114)
  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 8114)
  AUTHORS   Thole V, Vain P.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-DEC-2013) John Innes Centre, Norwich Research Park, 
            Norwich, Norfolk NR4 7UH, United Kingdom
REFERENCE   3  (bases 1 to 8114)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8114
                     /organism="Binary vector pCLEAN-G265"
                     /mol_type="other DNA"
                     /db_xref="taxon:1470146"
     misc_feature    1..2597
                     /note="derived from binary vector pCLEAN-G181 in INSD 
                     accession EU186083"
     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="NPTI"
                     /label=nptI
                     /note="kanamycin resistance protein"
                     /protein_id="AHL44326.1"
                     /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    2508..2532
                     /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    2533..2558
                     /note="spacer sequence separating left border No.1 and No.2
                     of the double left border sequence"
     misc_feature    2559..2583
                     /note="nopaline-type T-DNA left border sequence No.2 
                     belonging to a double left border sequence separated by a 
                     spacer sequence from left border No.1"
     regulatory      complement(2607..2859)
                     /regulatory_class="terminator"
                     /note="NOS-ter; poly(A)signal of nopaline synthase gene"
     terminator      2607..2859
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(2874..3425)
                     /codon_start=1
                     /gene="bar"
                     /product="phosphinothricin acetyl transferase"
                     /label=bar
                     /note="herbicide resistance protein"
                     /protein_id="AHL44327.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     gene            complement(2874..3425)
                     /gene="bar"
                     /label=bar
     intron          complement(3458..3991)
                     /note="adh1-intron 1; first intron of maize alcohol 
                     dehydrogenase 1 (adh1-1S)"
     regulatory      complement(4022..4443)
                     /regulatory_class="promoter"
                     /note="35S promoter from CaMV"
     promoter        complement(4022..4366)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     regulatory      complement(4466..4758)
                     /regulatory_class="terminator"
                     /note="SPA-ter; soybean-derived poly(A) signal"
     promoter        complement(4803..6799)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     intron          complement(4803..5816)
                     /note="ubi1-intron 1; modified first intron of maize 
                     ubiquitin-1"
     regulatory      complement(5817..6799)
                     /regulatory_class="promoter"
                     /note="ubi1-pro; promoter of maize ubiquitin-1"
     misc_feature    6849..6952
                     /note="UniBox2; synthetic unique sequence containing 
                     forward and reverse primer sequences"
     regulatory      6959..6983
                     /regulatory_class="other"
                     /note="nopaline-type T-DNA right border sequence"
     misc_feature    6959..6983
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    6984..7007
                     /label=right border overdrive sequence
                     /note="right border overdrive sequence"
     CDS             7211..8014
                     /codon_start=1
                     /gene="virGwt"
                     /product="VirG"
                     /label=virGwt
                     /note="wild-type"
                     /protein_id="AHL44328.1"
                     /translation="MIVHPSRENFSSAVNKGSDFRLKGEPLKHVLLVDDDVAMRHLIIE
                     YLTIHAFKVTAVADSTQFTRVLSSATVDVVVVDLNLGREDGLEIVRNLAAKSDIPIIII
                     SGDRLEETDKVVALELGASDFIAKPFSIREFLARIRVALRVRPNVVRSKDRRSFCFTDW
                     TLNLRQRRLMSEAGGEVKLTAGEFNLLLAFLEKPRDVLSREQLLIASRVRDEEVYDRSI
                     DVLILRLRRKLEADPSSPQLIKTARGAGYFFDADVQVSHGGTMAA"
     gene            7211..8014
                     /gene="virGwt"
                     /label=virGwt