LOCUS       Exported               14054 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pCaPVX440, complete sequence.
ACCESSION   KJ690769
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14054)
  AUTHORS   Wang Y, Cong QQ, Lan YF, Geng C, Li XD, Liang YC, Yang ZY, Zhu XP, 
            Li XD.
  TITLE     Development of new potato virus X-based vectors for gene 
            over-expression and gene silencing assay
  JOURNAL   Virus Res. 191, 62-69 (2014)
  PUBMED    25076104
REFERENCE   2  (bases 1 to 14054)
  AUTHORS   Wang Y, Cong QQ, Li XD.
  TITLE     Direct Submission
  JOURNAL   Submitted (10-APR-2014) Department of Plant Pathology, Shandong 
            Agricultural University, No. 61 Daizong street, Taian, Shandong 
            Province 271018, PR China
REFERENCE   3  (bases 1 to 14054)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14054
                     /organism="Expression vector pCaPVX440"
                     /mol_type="other DNA"
                     /db_xref="taxon:1530266"
     5UTR           1..84
                     /label=derived from PVX isolate 1985
                     /note="derived from PVX isolate 1985"
     CDS             85..4455
                     /codon_start=1
                     /gene="RdRp"
                     /product="RNA-dependent RNA polymerase"
                     /label=RdRp
                     /protein_id="AII77186.1"
                     /translation="MAKVREVYQSFTDSTTKTLIQDEAYRNIRPIMEKHKLSNPYAQTV
                     EAANDLEGFGIATNPYSIELHTHAAAKTIENKLLEVLGSILPQEPVTFMFLKPRKLNYM
                     RRNPRIKDIFHNVAIEPRDVARYPKETIIDKLTEITTDTAYISDTLHFLDPSYIVETFQ
                     NCPKLQTLYATLVLPVEAAFKMESTHPNIYSLKYFGDGFQYIPGNHGGGAYHHEFAHLQ
                     WLKVGKIKWRDPKDSFLGHLNYTTEQVEMHTVTVQLQESFAANHLYCIRRGDWLTPEVR
                     TFGQPDRYVIPPQIFLPKVHNCKKPILKKTMMQLFLYVRTVKVAKNCDIFAKVRQLIKS
                     SDLDKYSAVELVYLVSYMEFLADLQATTCFSDTLSGGLLTKTLAPVRAWIQEKKMQLFG
                     LEDYAKLVKAVDFHPVDFSFKVETWDFRFHPLQAWKAFRPREVSDVEEMESLFADGDLL
                     DCFTRMPAYAVNAEEDLAAIRKTPEMDVGQGIKEPAGDRNQYSNPAETFLNKLHRKHSR
                     EVKHQTTKKAKRLAEIQESMRAEGEAESNEMSGGMGAIPSNAELPSTSDARQELTLPTT
                     KSVPARWEDASLTDSSVEEEQVKLPGKEAVETAMQQVIEGLPWKHWIPQLNAVGFKALE
                     IQRDRNGTMIMPITEMVSGLEKEDFPEGTPKELARELLAMNRSPATIPLDLLRARDYGS
                     DVKNKRIGAITKTQATSWGEYLTGKIESLTERKVATCVIHGAGGSGKSHAIQKALREIG
                     KGSDITVVLPTNELRLDWSKKVPNTEPYMFKTYEKALIGGTGSIVIFDDYSKLPPGYIE
                     ALICFYSKIKLVILTGDSRQSVYHETAEDASIRHLGPATEYFSKYCRYYLNATHRNKKD
                     LANMLGVYSERTGVTEISMSAEFLEGIPTLVPSDEKRKLYMGTGRNDTFTYAGCQGLTK
                     PKVQIVLDHNTQVCSANVMYTALSRATDRIHFVNTSANSSAFWEKLDSTPYLKTFLSVV
                     REQALKEYEPAEVEPIREPEPQTHMCVENEESVLEEYKEELFEKFDREIHSESHGHSNC
                     VQTEDTTIQLFSHQQAKDETLLWATIDARLKTSNQETNFREFLSKKDIGDVLFLNYQKA
                     MGLPKERIPFSQEVWEACAHEVQSKYLSKSKCNLINGTVRQSPDFDENKIMVFLKSQWV
                     TKVEKLGLPKIKPGQTIAAFYQQTVMLFGTMARYMRWFRQAFQPKEVFINCETTPEDMS
                     AWALNNWNFGRPSLANDYTAFDQSQDGAMLQFEVLKAKHHCIPEEIIQAYIDIKTNAQI
                     FLGTLSIMRLTGEGPTFDANTECNIAYTHTKFDIPAGTAQVYAGDDSALDCVPEVKPSF
                     HRLEDKLLLKSKPVITQQKKGSWPEFCGWLITPKGVMKDPIKLHVSLKLAEAKGELKKC
                     QDSYEIDLSYAYDHKDSLHDLFDEKQCQAHTLTCRTLIKSGRGTVSLPRLRNFL"
     gene            85..4455
                     /gene="RdRp"
                     /label=RdRp
                     /note="derived from PVX isolate 1985"
     CDS             4486..5166
                     /codon_start=1
                     /gene="TGB1"
                     /product="triple gene block protein 1"
                     /label=TGB1
                     /protein_id="AII77188.1"
                     /translation="MDILIISLKSLGYSRTSKSLDSGPLVVHAVAGAGKSTALRKLILR
                     HPTFTVHTLGVPDKVSIRTRGIQKPGPIPEGNFPILDEYTLDNTTRDSYQALFADPYQA
                     PEFSLEPHFYLETSFRVPRKVADLIAGCGFDFETNSQEEGHLEITGIFKGPLLGKVIAI
                     DEESETTLSRHGVEFVKPCQVTGLEFKVVTIVSAAPIEEIGQSTAFYNAITRSKGLTYV
                     RAGT"
     gene            4486..5166
                     /gene="TGB1"
                     /label=TGB1
                     /note="derived from PVX isolate 1985"
     CDS             5147..5494
                     /codon_start=1
                     /gene="TGB2"
                     /product="triple gene block protein 2"
                     /label=TGB2
                     /protein_id="AII77187.1"
                     /translation="MSAQGHRLTAPVNSEKVYIVLGLSFALVSITFLLSRSNLPHVGDN
                     IHSLPHGGAYRDGTKAILYNSPNLGSRVSLHNGKNAAFAAVLLLTLLIYGSKYISQRNH
                     TCACGNNHSSH"
     gene            5147..5494
                     /gene="TGB2"
                     /label=TGB2
                     /note="derived from PVX isolate 1985"
     CDS             5427..5639
                     /codon_start=1
                     /gene="TGB3"
                     /product="triple gene block protein 3"
                     /label=TGB3
                     /protein_id="AII77189.1"
                     /translation="MEVNTYLNAIILVLVVTIIAVISTSLVRTEPCVIKITGESITVLA
                     CKLDAETIRAIADLKPLSVERLSFH"
     gene            5427..5639
                     /gene="TGB3"
                     /label=TGB3
                     /note="derived from PVX isolate 1985"
     RBS             5496..5504
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     misc_feature    5668..5691
                     /label=cloning site
                     /note="cloning site"
     regulatory      5692..5902
                     /regulatory_class="promoter"
                     /note="derived from tobacco mosaic virus coat protein 
                     subgenomic promoter"
     misc_feature    5903..5932
                     /label=multiple cloning site
                     /note="multiple cloning site"
     regulatory      5933..6014
                     /regulatory_class="promoter"
                     /note="derived from PVX isolate 1985"
     CDS             5997..6710
                     /codon_start=1
                     /gene="cp"
                     /product="coat protein"
                     /label=cp
                     /protein_id="AII77190.1"
                     /translation="MSSSASTTQATGSTTSTTTKTAGATPATASGLFTIPDGDFFSTAR
                     AIVASNAVATNEDLSKIEAIWKDMKVPTDTMAQAAWDLVRHCADVGSSAQTEMIDTGPY
                     SNGISRARLAAAIKEVCTLRQFCMKYAPVVWNWMLTNNSPPANWQAQGFKPEHKFAAFD
                     FFNGVTNPAAIMPKEGLIRPPSEAEMNAAQTAAFVKITKARAQSNDFASLDAAVTRGRI
                     TGTTTAEAVVTLPPP"
     gene            5997..6710
                     /gene="cp"
                     /label=cp
                     /note="derived from PVX isolate 1985"
     3UTR           6711..6811
                     /label=derived from PVX isolate 1985
                     /note="derived from PVX isolate 1985"
     misc_feature    6812..13618
                     /label=derived from binary vector pCambia0390
                     /note="derived from binary vector pCambia0390"
     CDS             6868..6885
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      6920..7172
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    7194..7218
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             8518..9147
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             9581..10648
                     /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      10714..10908
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    11252..11392
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(11578..12166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(12253..13047)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    13472..13496
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      13619..14054
                     /regulatory_class="promoter"
                     /note="CaMV 35S"
     promoter        13711..2
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"