LOCUS       Exported               10347 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector PCY, complete sequence.
ACCESSION   MG637043
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10347)
  AUTHORS   Cui T, Song Z.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-DEC-2017) Citrus Research Institute, Southwest 
            University, Xie ma, Beibei, Chongqing 400712, China
REFERENCE   2  (bases 1 to 10347)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..10347
                     /organism="Shuttle vector PCY"
                     /mol_type="other DNA"
                     /label=Ternary shuttle vector
                     /note="Ternary shuttle vector"
                     /db_xref="taxon:2282650"
     regulatory      1..767
                     /regulatory_class="promoter"
                     /note="derived from gene 2x35S"
     promoter        722..767
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     gene            771..1487
                     /gene="mgfp5"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
     CDS             771..1487
                     /codon_start=1
                     /gene="mgfp5"
                     /product="GFP with folding enhancement mutations"
                     /label=mgfp5
                     /note="suitable for expression in plants due to removal of 
                     a cryptic intron"
                     /translation="MSKGEELFTGVVPILVELDGDVNGYKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNFHNVYIMADKQKNGIKA
                     NFKTRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    1589..1841
                     /note="NOS terminator; nopaline synthase terminator and 
                     poly(A) signal"
     terminator      1589..1841
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     repeat_region   1863..1887
                     /label=right border repeat from nopaline C58 T-DNA
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    1863..1887
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3187..3816
                     /codon_start=1
                     /product="pVS1 StaA"
                     /label=pVS1 StaA
                     /note="stability protein from the Pseudomonas plasmid pVS1"
                     /protein_id="AXG50152.1"
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4211..5317
                     /codon_start=1
                     /product="pVS1 RepA"
                     /label=pVS1 RepA
                     /note="replication protein from the Pseudomonas plasmid 
                     pVS1"
                     /protein_id="AXG50153.1"
                     /translation="MALEPPSPRNRRDGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQR
                     IEAEARPGESWQAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSR
                     SIMDVAVFRLSKRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESM
                     NRYREGKGDKPGRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAK
                     NGRLVTVSEGEALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFL
                     IDPGIGRFLYRLARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYD
                     LKEEAGQAGPILVMRYRNLIEGEASAGS"
     CDS             4250..5317
                     /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      5383..5577
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1"
     misc_feature    5921..6061
                     /label=basis of mobility region from pBR322
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(6247..6835)
                     /direction=LEFT
                     /note="high copy number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6922..7716)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="KanR"
                     /label=aphA-3
                     /note="confers resistance to kanamycin"
                     /protein_id="AXG50154.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(6922..7716)
                     /gene="aphA-3"
                     /label=aphA-3
                     /note="aminoglycoside phosphotransferase"
     CDS             complement(8055..8729)
                     /codon_start=1
                     /gene="TRP1"
                     /product="TRP1"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /protein_id="AXG50155.1"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQLKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     gene            complement(8055..8729)
                     /gene="TRP1"
                     /label=TRP1
                     /note="derived from S. cerevisiae"
     regulatory      complement(8730..9010)
                     /regulatory_class="promoter"
                     /note="TRP1 promoter"
     promoter        complement(8730..9010)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     misc_feature    9269..9772
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     regulatory      9809..9913
                     /regulatory_class="promoter"
                     /note="AmpR promoter"
     promoter        9809..9913
                     /gene="bla"
                     /label=AmpR promoter
     repeat_region   10258..10282
                     /note="left border repeat from nopaline C58 T-DNA; LB T-DNA
                     repeat"
     misc_feature    10258..10282
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"