LOCUS       Exported               10970 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pFGUS2b, complete sequence.
ACCESSION   KC920578
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10970)
  AUTHORS   Zhang C, Chen ZJ.
  TITLE     Regulation of NAC2 expression by miR164c mediates fertilizer 
            application-induced delay of leaf senescence
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 10970)
  AUTHORS   Zhang C, Chen ZJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (18-APR-2013) Section of Molecular Cell & Developmental 
            Biology, The University of Texas at Austin, Austin, TX 78712, USA
REFERENCE   3  (bases 1 to 10970)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Vector NTI v. Suite 8
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10970
                     /organism="Binary vector pFGUS2b"
                     /mol_type="other DNA"
                     /db_xref="taxon:1473576"
     misc_feature    1..27
                     /note="T-Border (Left); left border repeat from C58 T-DNA"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      111..363
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     3UTR           complement(111..362)
                     /note="MAS polyA; polyA signal from mannopine synthase"
     CDS             complement(373..924)
                     /codon_start=1
                     /gene="BAR"
                     /product="BAR"
                     /label=BAR
                     /note="Phosphinothricin acetyltransferase (PAT), conferring
                     tolerance to the herbicide phosphinothricin (PPT)"
                     /protein_id="AHN10444.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     gene            complement(373..924)
                     /gene="BAR"
                     /label=BAR
     regulatory      complement(930..1310)
                     /regulatory_class="promoter"
                     /note="MAS promoter; mannopine synthase promoter"
     promoter        complement(930..1310)
                     /label=MAS promoter
                     /note="mannopine synthase promoter (Velten et al., 1984)"
     misc_feature    complement(1317..1592)
                     /note="NOS terminator sequence to insulate MAS promoter 
                     from the promoter to be cloned; or to be used as a polyA 
                     when a second transgene is inserted."
     terminator      1332..1584
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1593..1695
                     /note="Multiple Cloning Sites (MCS); MCS; synthetic linker 
                     sequence including 9 unique restriction sites"
     CDS             join(1696..1710,1901..3721)
                     /codon_start=1
                     /gene="gusA"
                     /product="GUS (beta-glucuronidase)"
                     /label=gusA
                     /protein_id="AHN10445.1"
                     /translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES
                     ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK
                     VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK
                     KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV
                     ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRS
                     VAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP
                     YAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA
                     IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD
                     AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT
                     LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK
                     GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            1696..3721
                     /gene="gusA"
                     /label=gusA
                     /note="GUS coding sequence amplified from pCAMBIA1381Z; 
                     without immunoTag; with confirmed sequence (10 nt 
                     corrected)"
     intron          1711..1900
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     CDS             1916..3721
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="RPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAIAV
                     PGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEVME
                     HQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHDFF
                     NYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQVV
                     ATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQFLI
                     NHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDWAD
                     EHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARDKN
                     HPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISDLF
                     DVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMYTD
                     MWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRKPK
                     SAAFLLQKRWTGMNFGEKPQQGGKQ"
     misc_feature    3728..1310
                     /label=sequence from pFGCGW INSD accession DQ231581.1
                     /note="sequence from pFGCGW INSD accession DQ231581.1"
     3UTR           3728..4447
                     /note="OCS polyA; polyA signal from octopine synthase"
     terminator      3738..4445
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     primer_bind     complement(4474..4490)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    4693..4717
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    4693..4716
                     /note="T-Border (Right); right border repeat from C58 
                     T-DNA"
     rep_origin      complement(5778..6778)
                     /direction=LEFT
                     /label=pVS1 replication origin
                     /note="pVS1 replication origin"
     CDS             6017..6646
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             7080..8147
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      8213..8407
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    8751..8891
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(9077..9665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      9181..9462
                     /label=pBR325 replication origin
                     /note="pBR325 replication origin"
     CDS             complement(9752..10546)
                     /codon_start=1
                     /gene="aadA"
                     /product="kanamycin (R)"
                     /label=aadA
                     /note="aadA (kanamycin resistance) gene amplified from 
                     pIG121Hm"
                     /protein_id="AHN10446.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(9752..10546)
                     /gene="aadA"
                     /label=aadA