LOCUS       Exported               14938 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLX-PPV, complete sequence.
ACCESSION   KY825150
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14938)
  AUTHORS   Pasin F, Bedoya LC, Bernabe-Orts JM, Gallo A, Simon-Mateo C, Orzaez 
            D, Garcia JA.
  TITLE     Multiple T-DNA Delivery to Plants Using Novel Mini Binary Vectors 
            with Compatible Replication Origins
  JOURNAL   ACS Synth Biol (2017) In press
  PUBMED    28657330
REFERENCE   2  (bases 1 to 14938)
  AUTHORS   Pasin F, Garcia JA.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAR-2017) Department of Plant Molecular Genetics, 
            Centro Nacional de Biotecnologia (CNB-CSIC), Darwin 3, Madrid, 
            Madrid 28049, Spain
REFERENCE   3  (bases 1 to 14938)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14938
                     /organism="Cloning vector pLX-PPV"
                     /mol_type="other DNA"
                     /db_xref="taxon:2016272"
     misc_feature    1..24
                     /label=assembly linker
                     /note="assembly linker"
     regulatory      25..72
                     /regulatory_class="terminator"
     misc_feature    133..157
                     /label=LB
                     /note="LB"
     misc_feature    180..210
                     /label=LB
                     /note="LB"
     misc_feature    184..208
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      440..1182
                     /regulatory_class="promoter"
                     /note="2x CaMV 35S"
     promoter        841..1185
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    join(1183..4106,4296..9947,10692..12000)
                     /label=Plum pox virus Rankovic
                     /note="Plum pox virus Rankovic"
     5UTR           1183..1328
     CDS             join(1329..4106,4296..11684)
                     /codon_start=1
                     /product="polyprotein"
                     /label=polyprotein
                     /protein_id="ASG92143.1"
                     /translation="MSTIVFGSFTCHLDAAIHQDNADRLAKAWTRPENHQVSNVHLLCR
                     RAAKSLINTYESATASAWKGLEEKLQPMFAKREFSKTVTKREGLRCFKESSEKFIEKKL
                     RKQYQEERERFQFLNGPDAIVNQISVDKCEASVRVPFPHIIEKPSFATPSMKKKVVFTK
                     VRMSEASLQLFMRRVAANAKANGQKVEIIGRKRVVGNYTTKSRLTYFRTHVRHLDGSKP
                     RYDLVLDEATKKILQLFANTSGFHHVHKKGEVTPGMSGFVVNPMNLSDPMQVYDTDLFI
                     VRGKHNSILVDSRCKVSKEQSNEIIHYSDPGKQFWDGFTNSFMQCKLRETDHQCTSDLD
                     VKECGYVAALVCQAIIPCGKITCLQCAQKYSYMSQQEIRDRFSTVIEQHEKTVMDNYPQ
                     FSHVLAFLKRYRELMRVENQNYEAFKDITHMIGERKEAPFSHLNKINELIIKGGMMSAQ
                     DYIEASDHLRELARYQKNRTENIRSGSIKAFRNKISSKAHVNMQLMCDNQLDTNGNFVW
                     GQREYHAKRFFRNYFDVIDVSEGYRRHIVRENPKGIRKLAIGNLVMSTNLAALRKQLLG
                     EECIHFEVSKECTSKRGENFVYQCCCVTHEDGTPLESEIISPTKNHLVVGNSGDSKYVD
                     LPTAKGGAMFIAKAGYCYINIFLAMLININEDEAKSFTKTVRDTLVPKLGTWPSMMDLA
                     TACHFLAILYPETRNAELPRILVDHEAKIFHVVDSFGSLSTGMHVLKANTINQLISFAS
                     DTLDSNMKTYLVGGLEVDKCDEFKNVKLLIRSIYKPQIMEQVLKEEPYLLLMSVLSPGV
                     LMALFNSGSLEKATQYWITRSHSLAAITSMLSSLAAKVSLASTLNAQMSVIDEHAAVLC
                     DSVFVGTKPYASYMMAVKTLERMKARTESDHTLNDLGFSVLRQATPHLVEKSYLQELEQ
                     AWKELSWSEKFSAILESQRWRKHIPKPFIPKDGADLGGRYDISVRSLLGNQYKRLRDVV
                     RRKRDDVVCYTHQSMGKLFCKAIGISTSFLPSTLKMFDMLIVFSLLLSIGATCNSMINE
                     HKHLKQLAADREDKKRFKRLQVLHTRLSEKVGCTPTADEFLEYVGGENPDLLKHAEDLI
                     GDGQVVVHQSKRDSQANLERVVAFVALVMMLFDSERSDGVYKILNKLKGIMGSVDQAVQ
                     HQSLDDIEDILDEKKLTVDFVLQSNEVAPTVPFDSTFEKWWTNQLETGNVIPHYRTEGH
                     FLEFTRENAAHIANEVMHGSHQDILIRGAVGSGKSTGLPFHLSKKGHVLLIEPTRPLAE
                     NVCKQLRGQPFNVNPTLRMRGMSTFGSTPITVMTSGYALHFLANNPTYLDNYKCIIFDE
                     CHVHDASAMAFRCLLSEYSYPGKILKVSATPPGHEVDFKTQKEVKVIVEESLSFQQFVS
                     NLGTGCNSDILKHGVNVLVYVASYNEVDTLSKLLTDRSFKVSKVDGRTMKIGNVEIPTS
                     GTQAKPHFVVATNIIENGVTLDIDVVVDFGLKVVPVLDIDNRLVRYTKKSISYGERIQR
                     LGRVGRNKPGAALRIGFTEKGLTQIPPIIATEAAFLCFTYGLPVMTNGVSTSLLAMCTV
                     KQARTMQQFELSPFYTVALVRFDGTMHQEIFRLLKSYRLRDSEVILNKLAIPNSNVCGW
                     MSVRDYKRQGCNLDLDENIRVPFYVKDIPETLHERIWQVVETHKSDAGFGRICSSSACK
                     IAYTLQTDIHSIPRTIKIIDALLEQERTKQAHFRAMTSQSCSSSNFSLSSITSAIRSKY
                     AKDHTEENIGVLQTAKSQLLEFKNLNIDPSYPELVRNFGALECVHHQTKEGVSKALQLK
                     GHWNKRLITRDATLMLGVLGGGAWMIFSYLRDSFKEEVVHQGFNRRQRQKLKFRQARDN
                     RMAREVYGDDSTMADYFGSAYSKKGKSKGKTRGMGTKTRKFVNMYGYDPTDYNFVRFVD
                     PLTGHTLDENPLMDINLVQEHFSQIRNDYIGDDKITMQHIMSNPGIVAYYIKDATQKAL
                     KVDLTPHNPLRVCDKTATIAGFPEREFELRQTGHPVFVEPNAIPKINEEGDEEVDHESK
                     SLFRGLRDYNPIASSICQLNNSSGARQSVMFGLGFGGLIVTNQHLFKRNDGELTIRSHH
                     GEFVVKDTKTLKLLPCKGRDIVIIRLPKDFPPFPKRLQFRTPTTEDRVCLIGSNFQTKS
                     ISSTMSETSATYPVDNSHFWKHWISTKDGHCGLPIVSTRDGSILGLHSLANSTNTQNFY
                     AAFPDNFETTYLSNQDNDNWIKQWRYNPDEVCWGSLQLKRDIPQSPFTICKLLTDLDGE
                     FVYTQSKTTHWLRDRLEGNLKAVGACPGQLVTKHVVKGKCTLFETYLLTHPEEHEFFRP
                     LMGAYQKSALNKDAYVKDLMKYSKPIVVGAVDCDQFERAVDVVISMLISKGFEECNYVT
                     DPDDIFSALNMKAAVGALYSGKKRDYFKNVSDQDKESFVRASCKRLFMGKKGVWNGSLK
                     AELRPKEKVEANKTRSFTAAPIDTLLGGKVCVDDFNNQFYSLNLHCPWSVGMTKFRGGW
                     DKLLRALPEGWIYCDADGSQFDSSLSPYLINAVLNIRLAFMEEWDIGEQMLSNLYTEIV
                     YTPIATPDGTIVKKFKGNNSGQPSTVVDNTLMVILAMTYSLLKLGYHPDTHDCICRYFV
                     NGDDLVLAVHPAYESIYDELQEHFSQLGLNYTFATKTENKEELWFMSHKGVLYDDMYIP
                     KLEPERIVSILEWDRSNEPIHRLEAICASMVEAWGYKELLREIRKFYSWVLEQAPYNAL
                     SKDGKAPYIAETALKKLYTDTEASETEIERYLEAFYDDFNDDGESNVVVHQAMVSKGEE
                     LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFT
                     YGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE
                     LKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQ
                     QNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKGTNV
                     VVHQADEREDEEEVDAGKPSVVTAPAATSPILQPPPVIQPAPRTTASMLNPIFTPATTQ
                     PATKPVSQVSGPQLQTFGTYGNEDASPSNSNALVNTNRDRDVDAGSVGTFTVPRLKAMT
                     SKLSLPKVKGKAIMNLNHLAHYSPAQVDLSNTRAPQSCFQTWYEGVKRDYDVTDDEMSI
                     ILNGLMVWCIENGTSPNINGMWVMMDGETQVEYPIKPLLDHAKPTFRQIMAHFSNVAEA
                     YIEKRNYEKAYMPRYGIQRNLTDYSLARYAFDFYEMTSTTPVRAREAHIQMKAAALRNV
                     QNRLFGLDGNVGTQEEDTERHTAGDVNRNMHNLLGVRGV"
     mat_peptide     1329..2252
                     /product="P1"
     mat_peptide     2253..3626
                     /product="HCPro"
     mat_peptide     join(3627..4106,4296..4865)
                     /product="P3"
     intron          4107..4295
     mat_peptide     4866..5021
                     /product="6K1"
     mat_peptide     5022..6926
                     /product="CI"
     mat_peptide     6927..7085
                     /product="6K2"
     mat_peptide     7086..8393
                     /product="NIa"
     mat_peptide     8394..9947
                     /product="NIb"
     misc_feature    9948..10691
                     /note="sGFP(S65T); GFP"
     CDS             9951..10667
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     mat_peptide     10692..11681
                     /product="CP"
     3UTR           11685..11901
     regulatory      12004..12261
                     /regulatory_class="terminator"
                     /note="nos"
     terminator      12014..12266
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    12304..12327
                     /label=RB
                     /note="RB"
     regulatory      complement(12392..12432)
                     /regulatory_class="terminator"
     misc_feature    12442..12465
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(12476..13291)
                     /codon_start=1
                     /gene="neo"
                     /product="neomycin phosphotransferase"
                     /label=neo
                     /note="kanamycin resistance protein; NptI"
                     /protein_id="ASG92144.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(12476..13291)
                     /gene="neo"
                     /label=neo
     misc_feature    13393..13416
                     /label=assembly linker
                     /note="assembly linker"
     CDS             complement(13417..14079)
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /protein_id="ASG92145.1"
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     gene            complement(13417..14079)
                     /gene="rep"
                     /label=rep
     rep_origin      14080..14851
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"