LOCUS       Exported               11005 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGR-PepMV-ON, complete sequence.
ACCESSION   KY031324
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11005)
  AUTHORS   Wu X, Wang A.
  TITLE     Evaluation the resistance of tomato varieties against Pepino mosaic 
            virus with an Ontario PepMV isolate
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11005)
  AUTHORS   Wu X, Wang A.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-OCT-2016) Agriculture and Agri-Food Canada, London 
            Research and Development Centre, 1391 Sandford Street, London, 
            Ontario N5V4T3, Canada
REFERENCE   3  (bases 1 to 11005)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..11005
                     /organism="Cloning vector pGR-PepMV-ON"
                     /mol_type="other DNA"
                     /country="Canada"
                     /note="derived from Pepino mosaic virus infectious clone 
                     Ontario-3 isolated from Solanum lycopersicum"
                     /db_xref="taxon:1919061"
     mobile_element  64..831
                     /mobile_element_type="insertion sequence:IS1"
                     /note="prokaryotic transposable element"
     rep_origin      1036..1583
                     /label=Rep_origin pSa
                     /note="Rep_origin pSa"
     rep_origin      1037..1472
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     misc_feature    1623..1647
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     protein_bind    1886..1907
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1922..1952
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1960..1976
                     /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     1984..2000
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2021..2039
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      2055..2489
                     /regulatory_class="promoter"
                     /note="CaMV 35S Promoter"
     promoter        2147..2492
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     5UTR           2490..2574
                     /label=PepMV 5 UTR
                     /note="PepMV 5 UTR"
     CDS             2575..6894
                     /codon_start=1
                     /gene="PepMV RdRp"
                     /product="RNA-dependent RNA polymerase"
                     /label=PepMV RdRp
                     /protein_id="APO36649.1"
                     /translation="MSRVRNTLEKIRDPQVQSSICEAAYQHVRPVLKESLINCPYALND
                     YEADTLENLGVTINPHAIQTHTHAAAKVVENRMLEIVGHHLPKDEKVTFIFLKRSKLRY
                     MRRAAVHKDVFVNHNIEPKDFFRYDEESTSTSFSVNTRIAYISDSLHFMEPADVTHLFD
                     RCQNLKTLMATVVLPVEAIHRQTSLFPAIYSINYNEEGFEYIPGSHGGGAYFHKYETLD
                     WLKYSRFIGHDPLTGLKYTITIQMVESLGANHLFLFQRGNFETPLYRTFQKNSFVTFPN
                     IFHPRHVNATKPMPRSRAIQLYLYVKSVNKVTQRDIFAKVRQLISTAELELYDPDELTH
                     VVNYFSYVSQLSSINDYDNMLKSSFFKKLVAPMQHDWRCMIEFFRGKSDFNQLLTALQW
                     KDFSYTIKTEELIITTHTAIGQAISGAANTYKERRQLTQLVKQGVISLADFQSEEPKIE
                     YTEFERETKPPVDCVTNYNNAVKNLGLSEPVDLPECSYARNSVPNNEISMAMTDADNFA
                     VINEIESLLSEEIAAPTLPALKNKTWASYASDTSSKKNEEIENIIAELEASRKVSNVQQ
                     TQHNYHISQCPTSLTAELPWKAWLPLLNAHGFKGDQIQHSPDGQIIQPIQDINNKTPRS
                     EYPSSIPADLVSTLRNIKRAVYAIPISHRRASAYSSDVKNNRTGKLLCAQSKEWKESFA
                     FKMQHEDIVKSGVVIHGCGGSGKSQALQNFLRTLGDNNDCCTVVVPTVELRNDWVNKLC
                     KLPMEHIKTFEKAMIQPGFPVVIFDDYTKLPPGYIEAYLFHHANTELFILTGDSRQSVY
                     HESNNEAYIASLDEAVAYYSNYCGFYLNATHRNVRSLANKLGVYSEKEGHLKITFASNA
                     LQKCKVPILVPSQMKKNAMQDIGHKAMTYAGCQGLTAPRVQILLDNHTQHCSDRVLYTC
                     LSRAVDSIHFINTGPNNSEFWDKLEATPYLKAFIDTYRDEKTEMLNSKPADDSPVEPEA
                     PATHFPVSNGNNLEKLASTLPEKFAREIYDKHHGYSNTIQTENPIVQLFQHQQAKDETL
                     FWATIEARLSITTPDANLREFTLKKDVGDILFFNYHSAMCLPADPVDFEPRTWEICAAE
                     VKNTYLAKPMANLINAASRQSPDFEPNKISLFLKSQWVKKVEKLGAIKSKPGQTIAAFM
                     QQTVMLYGTMARYLRKMRQRFQPKHIFINCETTTDDLNNFVLNGWNFNRTAQTNDFTAF
                     DQSQDGAMLQFEVMKAKFFNIPADVIEGYINIKLNAKIFLGTLSIMRLSGEGPTFDANT
                     ECSIAYTATRYHLSSAVKQVYAGDDMALDGVVMEKPSFKKLQSKLKLTSKTLFPKQVKG
                     DYAEFCGWTFTPGGIIKNPLKMHASIMLQEAIGNLHTAARSYAIDMKHSYQMGDKLHEY
                     LTPDEAEQHFLAVRKLHKLHQGEAMRLGEKSPPKATH"
     gene            2575..6894
                     /gene="PepMV RdRp"
                     /label=PepMV RdRp
                     /note="PepMVgp1"
     CDS             6919..7623
                     /codon_start=1
                     /gene="PepMV TGB1"
                     /product="TGBp1"
                     /label=PepMV TGB1
                     /note="movement protein"
                     /protein_id="APO36650.1"
                     /translation="MERSTLINLLQLHHFEPKLSVEGVIVVHGIAGTGKTTLLRTLFSA
                     YPNLVIGSPRPCYLDKANKISQVCLSCFPNTLCDIVDEYHLLESYQEPTLALFGDPCQC
                     TFIGRLRIPHYTSFRTHRFGKSTAELLNKLFQLQIVSVKQEDDIVEFFDPFQVDPTENI
                     SASEEEVLEFVSDQVVTTSSEELAGLEFTETTFYCTTLAAAVTENPARTFISLTRHTQR
                     LTIGELNARIDS"
     gene            6919..7623
                     /gene="PepMV TGB1"
                     /label=PepMV TGB1
                     /note="PepMVgp2"
     CDS             7604..7975
                     /codon_start=1
                     /gene="PepMV TGB2"
                     /product="TGBp2"
                     /label=PepMV TGB2
                     /note="movement protein"
                     /protein_id="APO36652.1"
                     /translation="MPGLTPRADLTDTYKIIAIAFLLSACIYFQNSHYQPVAGDNLHRL
                     PFGGQYQDGTKKISYFPQQQSYFHSGNKLNVLILIFILTLGIVLTNKFSFSVSRTTHQH
                     SCYNTHSAANTTPPLSGHH"
     gene            7604..7975
                     /gene="PepMV TGB2"
                     /label=PepMV TGB2
                     /note="PepMVgp3"
     CDS             7827..8075
                     /codon_start=1
                     /gene="PepMV TGB3"
                     /product="TGBp3"
                     /label=PepMV TGB3
                     /note="movement protein"
                     /protein_id="APO36653.1"
                     /translation="MSSYSSSFLHWVSSSPINLVLALAVLLTSILATTHILQQTQHHHC
                     QVIIDGAAIVVTNCENTPEVLKAINFAPWNGLSFPNV"
     gene            7827..8075
                     /gene="PepMV TGB3"
                     /label=PepMV TGB3
                     /note="PepMVgp4"
     CDS             8122..8835
                     /codon_start=1
                     /gene="PepMV CP"
                     /product="CP"
                     /label=PepMV CP
                     /note="coat protein"
                     /protein_id="APO36651.1"
                     /translation="MADNTPVAATSGSPPTAQDAGARAPADFSNPNTAPSLSDLKKVKY
                     VSTVTSVATPTEIEALGKIFTAMGLAANETGPAMWDLARAYADVQSSKSAQLIGATPSN
                     PALSRRALAAQFDRINITPRQFCMYFAKVVWNILLDSNIPPANWAKLGYQEDTKFAAFD
                     FFDGVTNPASLQPADGLIRQPNEKELAAHSVAKYGALARQKISTGNYITTLGEVTRGHM
                     GGANTMYAIDAPPEL"
     gene            8122..8835
                     /gene="PepMV CP"
                     /label=PepMV CP
                     /note="PepMVgp5"
     3UTR           8836..8904
                     /label=PepMV 3 UTR
                     /note="PepMV 3 UTR"
     regulatory      8949..9202
                     /regulatory_class="terminator"
                     /note="NOS terminator"
     terminator      8949..9201
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(9210..9226)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    9382..9404
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      9429..10083
                     /label=E.coli rep_origin colE1
                     /note="E.coli rep_origin colE1"
     rep_origin      complement(9471..10059)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(10160..10975)
                     /codon_start=1
                     /product="kanmycin"
                     /label=kanmycin
                     /note="kanmycin resistance in bacteria"
                     /protein_id="APO36654.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"