LOCUS       Exported               12184 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Plant binary expression vector pCaS1, complete sequence.
ACCESSION   KP836314
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12184)
  AUTHORS   Toosi S, Shokouhifar F, Malekzadeh Shafaroudi S, Bagheri A.
  TITLE     Functional Analysis of Fol-SIX1 in Resistant and Susceptible Melon 
            Lines
  JOURNAL   Zist Fanavarii Giyahani Zirai 7 (4), 35-47 (2015)
REFERENCE   2  (bases 1 to 12184)
  AUTHORS   Toosi S, Shokouhifar F, Malekzadeh Shafaroudi S, Bagheri A.
  TITLE     Direct Submission
  JOURNAL   Submitted (21-FEB-2015) Reserarch Center for Plant Sciences, 
            Ferdowsi University of Mashhad, Mashhad, Khorasan Razavi 91775-1653,
            Iran
REFERENCE   3  (bases 1 to 12184)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: seqman v. 7.0.0
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..12184
                     /organism="Plant binary expression vector pCaS1"
                     /mol_type="other DNA"
                     /db_xref="taxon:1630126"
     misc_feature    1..26
                     /note="T-DNA left border; RB"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      103..277
                     /regulatory_class="polyA_signal_sequence"
                     /note="CaMV 35S poly A"
     polyA_signal    103..277
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(284..835)
                     /codon_start=1
                     /gene="Bar"
                     /product="phosphinothricin N-acetyltransferase"
                     /label=Bar
                     /protein_id="AJW67225.1"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     gene            complement(284..835)
                     /gene="Bar"
                     /label=Bar
     regulatory      complement(848..1655)
                     /regulatory_class="promoter"
                     /note="CaMV 35S"
     promoter        complement(880..1557)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    1748..1769
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1784..1814
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1822..1838
                     /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     1846..1862
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1858..2091
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEC"
     misc_feature    1872..1928
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1932..1948)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2325..2670
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             2687..3541
                     /codon_start=1
                     /gene="Fol-SIX1"
                     /product="secreted in xylem 1 protein"
                     /label=Fol-SIX1
                     /protein_id="AJW67224.1"
                     /translation="MAPYSMVLLGALSILGFGAYAQEAAVREPQIFFNLTYTEYLDKVA
                     ASHGSPPDKSDLPWNDTMGSFPGNETDDGVQTETGSSLSRRGHIVNLRKREPFGEESRN
                     DRVTQDMLQALHDLCVERFGTGYRAVSGLCYTDRRATRKIECNKPSVRERDRSVTRACP
                     EGQECTTFNAYNFRNRHHQVTFPVCGPRIEVKDRHDIGIHTEWQGTWYPESPKSPGTYD
                     YFAQMAGTLNGYFGYDGVYSDGYKTSSHGYGHSWSCINCPRGKVTITNTYRATWAFGYT
                     SPH"
     gene            2687..3541
                     /gene="Fol-SIX1"
                     /label=Fol-SIX1
     CDS             join(3552..3566,3757..5604)
                     /codon_start=1
                     /gene="GUS"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /protein_id="AJW67226.1"
                     /translation="MVDLRNRRLVRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWES
                     ALQESRAIAVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGK
                     VWVNNQEVMEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGK
                     KKQSYFHDFFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSV
                     ELRDADQQVVATGQGTSGTLQVVNPHLWQPGEGYLYELEVTAKSQTESDIYPLRVGIRS
                     VAVKGQQFLINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYP
                     YAEEMLDWADEHGIVVIDETAAVGFQLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQA
                     IKELIARDKNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCD
                     AHTDTISDLFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDT
                     LAGLHSMYTDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKK
                     GIFTRDRKPKSAAFLLQKRWTGMNFGEKPQQGGKQASHHHHHHV"
     gene            3552..5604
                     /gene="GUS"
                     /label=GUS
     CDS             join(3552..3566,3757..5574)
                     /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"
     intron          3567..3756
                     /gene="GUS"
                     /note="derived from catalase"
     CDS             5581..5598
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      5633..5885
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      5636..5888
                     /regulatory_class="polyA_signal_sequence"
                     /note="Nos terminator"
     misc_feature    5907..5931
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    5926..5951
                     /note="T-DNA right border; RB"
     CDS             7231..7860
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             8294..9361
                     /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      9427..9621
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    9965..10105
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(10291..10879)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10966..11760)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"