LOCUS       Exported                6873 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pBrachyTAG-G220, complete sequence.
ACCESSION   HQ702996
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6873)
  AUTHORS   Thole V, Worland B, Vain P.
  TITLE     pBrachyTAG vector system: New tools for DNA tagging and trapping in 
            grasses
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6873)
  AUTHORS   Thole V, Vain P.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-DEC-2010) Crop Genetics, John Innes Centre, Norwich 
            Research Park, Colney Lane, Norwich, Norfolk NR4 7UH, United Kingdom
REFERENCE   3  (bases 1 to 6873)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6873
                     /organism="Binary vector pBrachyTAG-G220"
                     /mol_type="other DNA"
                     /db_xref="taxon:992201"
     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="ADZ04462.1"
                     /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 border sequence 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"
     misc_feature    2713..4652
                     /note="derived from binary vector pVec8-GFP in INSD 
                     accession FJ949107"
     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="ADZ04463.1"
                     /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 the 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"
     CDS             complement(4979..5698)
                     /codon_start=1
                     /gene="sgfp(S65T)"
                     /product="sGFP(S65T)"
                     /label=sgfp(S65T)
                     /note="synthetic green fluorescent protein"
                     /protein_id="ADZ04464.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     gene            complement(4979..5698)
                     /gene="sgfp(S65T)"
                     /label=sgfp(S65T)
     misc_feature    5718..5742
                     /label=nopaline-type T-DNA right border sequence
                     /note="nopaline-type T-DNA right border sequence"
     misc_feature    5743..5766
                     /label=right border overdrive sequence
                     /note="right border overdrive sequence"
     CDS             5970..6773
                     /codon_start=1
                     /gene="virGwt"
                     /product="virG"
                     /label=virGwt
                     /note="wild-type virG"
                     /protein_id="ADZ04465.1"
                     /translation="MIVHPSRENFSSAVNKGSDFRLKGEPLKHVLLVDDDVAMRHLIIE
                     YLTIHAFKVTAVADSTQFTRVLSSATVDVVVVDLNLGREDGLEIVRNLAAKSDIPIIII
                     SGDRLEETDKVVALELGASDFIAKPFSIREFLARIRVALRVRPNVVRSKDRRSFCFTDW
                     TLNLRQRRLMSEAGGEVKLTAGEFNLLLAFLEKPRDVLSREQLLIASRVRDEEVYDRSI
                     DVLILRLRRKLEADPSSPQLIKTARGAGYFFDADVQVSHGGTMAA"
     gene            5970..6773
                     /gene="virGwt"
                     /label=virGwt