LOCUS       Exported               14317 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Binary vector pVCPGUS(I)23010, complete sequence.
ACCESSION   JQ436738
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14317)
  AUTHORS   Singh A, Buttar BS, Gupta A, Khurana JP, Chaudhary VK.
  TITLE     Direct Submission
  JOURNAL   Submitted (23-JAN-2012) Department of Biochemistry, Department of 
            Microbiology and Department of Plant Molecular Biology, University 
            of Delhi South Campus, New Delhi, Delhi 110021, India
REFERENCE   2  (bases 1 to 14317)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14317
                     /organism="Binary vector pVCPGUS(I)23010"
                     /mol_type="other DNA"
                     /db_xref="taxon:1179823"
     regulatory      complement(29..113)
                     /regulatory_class="terminator"
                     /note="T7 terminator"
     terminator      29..76
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      complement(135..405)
                     /regulatory_class="polyA_signal_sequence"
                     /note="nopaline synthase (nos) 3 UTR"
     terminator      143..395
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(join(412..2307,2498..2512))
                     /codon_start=1
                     /product="GUS"
                     /label=GUS
                     /note="contains c-myc and 8 histidine tag"
                     /protein_id="AFI93667.1"
                     /translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES
                     ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK
                     VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK
                     KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV
                     ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRS
                     VAVKGEQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP
                     YAEEMLDWADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA
                     IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD
                     AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT
                     LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK
                     GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQASGEQKLISEEDLNSGGHHHHHHH
                     H"
     CDS             complement(415..438)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             complement(451..480)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             complement(490..2292)
                     /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"
     intron          complement(2308..2497)
                     /note="castorbean catalase intron"
     5UTR           complement(2536..2572)
                     /label=derived from alfalfa mosaic virus
                     /note="derived from alfalfa mosaic virus"
     promoter        complement(2577..2595)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      complement(2578..2595)
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     regulatory      2607..3138
                     /regulatory_class="promoter"
                     /note="CaMV35S; 35S promoter from CaMV"
     promoter        2790..3135
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             3152..5806
                     /codon_start=1
                     /product="T7 RNA polymerase"
                     /label=T7 RNA polymerase
                     /note="contains nuclear localization signal (NLS) from SV40
                     large T antigen"
                     /protein_id="AFI93668.1"
                     /translation="MEPPKKKRKVEFSDIELAAIPFNTLADHYGERLAREQLALEHESY
                     EMGEARFRKMFERQLKAGEVADNAAAKPLITTLLPKMIARINDWFEEVKAKRGKRPTAF
                     QFLQEIKPEAVAYITIKTTLACLTSADNTTVQAVASAIGRAIEDEARFGRIRDLEAKHF
                     KKNVEEQLNKRVGHVYKKAFMQVVEADMLSKGLLGGEAWSSWHKEDSIHVGVRCIEMLI
                     ESTGMVSLHRQNAGVVGQDSETIELAPEYAEAIATRAGALAGISPMFQPCVVPPKPWTG
                     ITGGGYWANGRRPLALVRTHSKKALMRYEDVYMPEVYKAINIAQNTAWKINKKVLAVAN
                     VITKWKHCPVEDIPAIEREELPMKPEDIDMNPEALTAWKRAAAAVYRKDKARKSRRISL
                     EFMLEQANKFANHKAIWFPYNMDWRGRVYAVSMFNPQGNDMTKGLLTLAKGKPIGKEGY
                     YWLKIHGANCAGVDKVPFPERIKFIEENHENIMACAKSPLENTWWAEQDSPFCFLAFCF
                     EYAGVQHHGLSYNCSLPLAFDGSCSGIQHFSAMLRDEVGGRAVNLLPSETVQDIYGIVA
                     KKVNEILQADAINGTDNEVVTVTDENTGEISEKVKLGTKALAGQWLAYGVTRSVTKRSV
                     MTLAYGSKEFGFRQQVLEDTIQPAIDSGKGLMFTQPNQAAGYMAKLIWESVSVTVVAAV
                     EAMNWLKSAAKLLAAEVKDKKTGEILRKRCAVHWVTPDGFPVWQEYKKPIQTRLNLMFL
                     GQFRLQPTINTNKDSEIDAHKQESGIAPNFVHSQDGSHLRKTVVWAHEKYGIESFALIH
                     DSFGTIPADAANLFKAVRETMVDTYESCDVLADFYDQFADQLHESQLDKMPALPAKGNL
                     NLRDILESDFAFA"
     CDS             3161..3181
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      5830..6082
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     3UTR           5833..6085
                     /label=nopaline synthase (nos) 3 UTR
                     /note="nopaline synthase (nos) 3 UTR"
     misc_feature    6104..6128
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    6123..6148
                     /label=right border repeat from pCAMBIA-2301
                     /note="right border repeat from pCAMBIA-2301"
     misc_feature    6981..7981
                     /label=STA region from pVS1 plasmid
                     /note="STA region from pVS1 plasmid"
     CDS             7428..8057
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8486..9559
                     /codon_start=1
                     /product="RepA"
                     /label=RepA
                     /note="derived from pVS1"
                     /protein_id="AFI93669.1"
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELVDWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
     rep_origin      8783..9783
                     /note="pVS1-REP; replication origin from pVS1"
     rep_origin      9625..9819
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    10163..10303
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    10193..10453
                     /label=bom site of pBR322 from pCAMBIA-2301
                     /note="bom site of pBR322 from pCAMBIA-2301"
     rep_origin      complement(10489..11077)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      10593..10873
                     /label=pBR322 origin of replication from pCAMBIA-2301
                     /note="pBR322 origin of replication from pCAMBIA-2301"
     CDS             complement(11164..11958)
                     /codon_start=1
                     /product="AadA"
                     /label=AadA
                     /note="kanamycin resistance protein; derived from 
                     pCAMBIA-2301"
                     /protein_id="AFI93670.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    12383..12408
                     /label=left border repeat from pCAMBIA-2301
                     /note="left border repeat from pCAMBIA-2301"
     misc_feature    12383..12407
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     3UTR           complement(12475..12678)
                     /label=CaMV 3 UTR from pCAMBIA-2301
                     /note="CaMV 3 UTR from pCAMBIA-2301"
     polyA_signal    12485..12659
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(12716..13513)
                     /codon_start=1
                     /product="NptII"
                     /label=NptII
                     /note="kanamycin resistance protein; derived from 
                     pCAMBIA-2301"
                     /protein_id="AFI93671.1"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(13544..14317)
                     /regulatory_class="promoter"
                     /note="CaMV35S2; CaMV 35S promoter; duplicated from 
                     pCAMBIA-2301"
     promoter        complement(13576..14253)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"