LOCUS       Exported               11633 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCAMBIA-2301, complete sequence.
ACCESSION   AF234316
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11633)
  AUTHORS   Hajdukiewicz P, Svab Z, Maliga P.
  TITLE     The small, versatile pPZP family of Agrobacterium binary vectors for
            plant transformation
  JOURNAL   Plant Mol. Biol. 25 (6), 989-994 (1994)
  PUBMED    7919218
REFERENCE   2  (bases 1 to 11633)
  AUTHORS   Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi
            KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet 
            I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A, 
            Jefferson RA.
  TITLE     A comprehensive set of modular vectors for advanced manipulations 
            and efficient transformation of plants
  JOURNAL   Unpublished
REFERENCE   3  (bases 1 to 11633)
  AUTHORS   Roberts C, Rajagopal S, Smith LM, Nguyen TA, Yang W, Nugrohu S, Ravi
            KS, Vijayachandra K, Harcourt RL, Dransfield L, Desamero N, Slamet 
            I, Hadjukiewicz P, Svab Z, Maliga P, Mayer JE, Keese PK, Kilian A, 
            Jefferson RA.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-FEB-2000) CAMBIA, Clunies Ross St, Black Mountain / 
            GPO Box 3200, Canberra, ACT 2601, Australia
REFERENCE   4  (bases 1 to 11633)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11633
                     /organism="Binary vector pCAMBIA-2301"
                     /mol_type="other DNA"
                     /db_xref="taxon:118408"
     CDS             join(2..16,207..2054)
                     /codon_start=1
                     /product="GUSA, hexaHis tagged"
                     /label=GUSA, hexaHis tagged
                     /protein_id="AAF65404.1"
                     /translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES
                     ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK
                     VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK
                     KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV
                     ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELEVTAKSQTESDIYPLRVGIRS
                     VAVKGQQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP
                     YAEEMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA
                     IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD
                     AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT
                     LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK
                     GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQASHHHHHHV"
     CDS             join(2..16,207..2024)
                     /codon_start=1
                     /product="beta-glucuronidase"
                     /label=GUS
                     /note="This version of the gusA gene has a 5 extension 
                     with a catalase intron to ensure expression in plants but 
                     not bacteria."
                     /translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES
                     ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK
                     VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK
                     KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV
                     ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELEVTAKSQTESDIYPLRVGIRS
                     VAVKGQQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP
                     YAEEMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA
                     IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD
                     AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT
                     LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK
                     GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     exon            2..16
                     /note="synthetic first gus exon (modular)"
     intron          17..206
                     /note="Castorbean catalase intron; modified"
     exon            207..2054
                     /note="second gusA exon, His6"
     CDS             2031..2048
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2083..2335
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2086..2338
                     /label=nos (nopaline synthase) 3UTR (polyA signal)
                     /note="nos (nopaline synthase) 3UTR (polyA signal)"
     misc_feature    2357..2381
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    2376..2401
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"
     misc_feature    complement(3442..4442)
                     /label=STA region from pVS1 plasmid
                     /note="STA region from pVS1 plasmid"
     CDS             3681..4310
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4744..5811
                     /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      complement(5035..6035)
                     /direction=LEFT
                     /note="pVS1-REP; replication origin from pVS1"
     rep_origin      5877..6071
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6415..6555
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(6445..6705)
                     /label=bom site from pBR322
                     /note="bom site from pBR322"
     rep_origin      complement(6741..7329)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(6845..7125)
                     /direction=LEFT
                     /label=pBR322 origin of replication
                     /note="pBR322 origin of replication"
     CDS             complement(7416..8210)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=aminoglycoside phosphotransferase
                     /note="aadA (kanamycin resistance) gene amplified from 
                     pIG121Hm"
                     /protein_id="AAF65402.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    8635..8660
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    8635..8659
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    8727..8930
                     /label=CaMV 3UTR (polyA signal)
                     /note="CaMV 3UTR (polyA signal)"
     polyA_signal    8737..8911
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(8968..9765)
                     /codon_start=1
                     /product="neomycin phosphotransferase"
                     /label=neomycin phosphotransferase
                     /note="nptII (kanamycin resistance) gene"
                     /protein_id="AAF65403.1"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      complement(9796..10576)
                     /regulatory_class="promoter"
                     /note="CaMV35S2; CaMV 35S promoter, duplicated"
     promoter        complement(9828..10505)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    10696..10717
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     regulatory      10724..10778
                     /regulatory_class="promoter"
                     /note="PlacZ; lacZ promoter"
     promoter        10732..10762
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10770..10786
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     10794..10810
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             10806..11039
                     /codon_start=1
                     /product="LacZ alpha fragment"
                     /label=LacZ alpha fragment
                     /protein_id="AAF65401.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     misc_feature    10820..10876
                     /note="pUC18 MCS; polylinker"
     primer_bind     complement(10880..10896)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      11090..11627
                     /regulatory_class="promoter"
                     /note="CaMV35S; 35S promoter from CaMV"
     promoter        11273..11618
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"