LOCUS       Exported                9085 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pBrachyTAG-G229, complete sequence.
ACCESSION   HQ711853
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9085)
  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  JOURNAL   Unpublished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 9085)
  AUTHORS   Thole V, Vain P.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-DEC-2010) Crop Genetics, John Innes Centre, Norwich 
            Research Park, Colney Lane, Norwich, Norfolk NR4 7UH, United Kingdom
REFERENCE   3  (bases 1 to 9085)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9085
                     /organism="Binary vector pBrachyTAG-G229"
                     /mol_type="other DNA"
                     /db_xref="taxon:992203"
     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="ADZ04466.1"
                     /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"
     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="ADZ04467.1"
                     /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"
     CDS             complement(4971..5765)
                     /codon_start=1
                     /gene="sgfp(S65T)-ER"
                     /product="sGFP(S65T)-ER"
                     /label=sgfp(S65T)-ER
                     /note="modified synthetic green fluorescent protein; 
                     contains sequences that code for signal peptides that 
                     result in retention of the protein in the plant endoplasmic
                     reticulum (ER)"
                     /protein_id="ADZ04469.1"
                     /translation="MKTNLFLFLIFSLLLSLSSAEFVSKGEELFTGVVPILVELDGDVN
                     GHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHD
                     FFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKL
                     EYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY
                     LSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKHDEL"
     gene            complement(4971..5765)
                     /gene="sgfp(S65T)-ER"
                     /label=sgfp(S65T)-ER
     regulatory      complement(4974..4985)
                     /regulatory_class="other"
                     /gene="sgfp(S65T)-ER"
                     /note="C-terminal HDEL signal sequence for protein 
                     secretion and retention in the ER"
     CDS             complement(4986..5699)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     regulatory      complement(5700..5765)
                     /regulatory_class="other"
                     /gene="sgfp(S65T)-ER"
                     /note="N-terminal ER signal peptide sequence from 
                     Arabidopsis thaliana basic chitinase"
     promoter        complement(5774..7770)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     intron          complement(5774..6787)
                     /note="ubi1-intron 1; modified first intron of maize 
                     ubiquitin-1 gene"
     regulatory      complement(6788..7770)
                     /regulatory_class="promoter"
                     /note="ubi1-pro; promoter of maize ubiquitin-1 gene"
     misc_feature    7820..7923
                     /note="UniBox2; synthetic unique sequence containing 
                     forward and reverse primer sequences"
     misc_feature    7930..7954
                     /label=nopaline-type T-DNA right border sequence
                     /note="nopaline-type T-DNA right border sequence"
     misc_feature    7955..7978
                     /label=right border overdrive sequence
                     /note="right border overdrive sequence"
     CDS             8182..8985
                     /codon_start=1
                     /gene="virGwt"
                     /product="virG"
                     /label=virGwt
                     /note="wild-type virG"
                     /protein_id="ADZ04468.1"
                     /translation="MIVHPSRENFSSAVNKGSDFRLKGEPLKHVLLVDDDVAMRHLIIE
                     YLTIHAFKVTAVADSTQFTRVLSSATVDVVVVDLNLGREDGLEIVRNLAAKSDIPIIII
                     SGDRLEETDKVVALELGASDFIAKPFSIREFLARIRVALRVRPNVVRSKDRRSFCFTDW
                     TLNLRQRRLMSEAGGEVKLTAGEFNLLLAFLEKPRDVLSREQLLIASRVRDEEVYDRSI
                     DVLILRLRRKLEADPSSPQLIKTARGAGYFFDADVQVSHGGTMAA"
     gene            8182..8985
                     /gene="virGwt"
                     /label=virGwt