LOCUS       Exported                8418 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pBrachyTAG-G227, complete sequence.
ACCESSION   JN153032
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8418)
  AUTHORS   Thole V, Worland B, Vain P.
  TITLE     pBrachyTAG vector system: New tools for T-DNA tagging and trapping 
            in grasses
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8418)
  AUTHORS   Thole V, Vain P.
  TITLE     Direct Submission
  JOURNAL   Submitted (17-JUN-2011) Department of Crop Genetics, John Innes 
            Centre, Norwich Research Park, Colney Lane, Norwich, Norfolk NR4 
            7UH, United Kingdom
REFERENCE   3  (bases 1 to 8418)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8418
                     /organism="Binary vector pBrachyTAG-G227"
                     /mol_type="other DNA"
                     /db_xref="taxon:1109410"
     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_id="AET79510.1"
                     /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 (Thole et al. (2007) 
                     Plant Physiol. 145 (4), 1211-1219)"
     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 No.2 belonging to a 
                     double left border sequence separated by a spacer sequence 
                     from left border No.1"
     misc_feature    2598..2693
                     /note="UniBox1; synthetic unique sequence containing 
                     forward and reverse primer sequences"
     regulatory      complement(2713..2965)
                     /regulatory_class="terminator"
                     /note="NOS-ter; poly(A) signal of nopaline synthase gene"
     terminator      2713..2965
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(join(3006..3692,3883..4221))
                     /codon_start=1
                     /gene="hptI"
                     /product="HPTI"
                     /label=hptI
                     /note="hygromycin phosphotransferase"
                     /protein_id="AET79511.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     gene            complement(3006..4221)
                     /gene="hptI"
                     /label=hptI
     intron          complement(3693..3882)
                     /gene="hptI"
                     /note="CAT-1 intron; modified first intron from bean 
                     catalase-1 gene"
     regulatory      complement(4231..4652)
                     /regulatory_class="promoter"
                     /note="35S promoter from CaMV"
     promoter        complement(4231..4575)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     regulatory      complement(4672..4964)
                     /regulatory_class="terminator"
                     /note="SPA-ter; soybean-derived poly(A) signal"
     misc_feature    4965..5106
                     /note="spacer sequence (containing non-useable ER targeting
                     signal sequences) to be replaced with gene-of-interest 
                     using unique cloning sites KpnI (nt 4965-4970) and HindIII 
                     (nt 5101-5106)"
     promoter        complement(5107..7103)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     intron          complement(5107..6120)
                     /note="ubi1-intron 1; modified first intron of maize 
                     ubiquitin-1 gene"
     regulatory      complement(6121..7103)
                     /regulatory_class="promoter"
                     /note="ubi1-pro; promoter of maize ubiquitin-1 gene"
     misc_feature    7153..7256
                     /note="UniBox2; synthetic unique sequence containing 
                     forward and reverse primer sequences"
     misc_feature    7263..7287
                     /label=nopaline-type T-DNA right border sequence
                     /note="nopaline-type T-DNA right border sequence"
     misc_feature    7288..7311
                     /label=right border overdrive sequence
                     /note="right border overdrive sequence"
     CDS             7515..8318
                     /codon_start=1
                     /gene="virGwt"
                     /product="VirG"
                     /label=virGwt
                     /note="wild-type virG"
                     /protein_id="AET79512.1"
                     /translation="MIVHPSRENFSSAVNKGSDFRLKGEPLKHVLLVDDDVAMRHLIIE
                     YLTIHAFKVTAVADSTQFTRVLSSATVDVVVVDLNLGREDGLEIVRNLAAKSDIPIIII
                     SGDRLEETDKVVALELGASDFIAKPFSIREFLARIRVALRVRPNVVRSKDRRSFCFTDW
                     TLNLRQRRLMSEAGGEVKLTAGEFNLLLAFLEKPRDVLSREQLLIASRVRDEEVYDRSI
                     DVLILRLRRKLEADPSSPQLIKTARGAGYFFDADVQVSHGGTMAA"
     gene            7515..8318
                     /gene="virGwt"
                     /label=virGwt