LOCUS       sequence-342138-        8398 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  This is a binary plasmid that can be transferred into Agrobacterium 
            and used to transiently express the protein encoded by the insert in
            plants by agro-infiltration..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8398)
  AUTHORS   Derevnina L, Contreras MP, Adachi H, Upson J, Vergara Cruces A, Xie 
            R, Sklenar J, Menke FLH, Mugford ST, MacLean D, Ma W, Hogenhout SA, 
            Goverse A, Maqbool A, Wu CH, Kamoun S
  TITLE     Plant pathogens convergently evolved to counteract redundant nodes 
            of an NLR immune receptor network.
  JOURNAL   PLoS Biol. 2021 Aug 23;19(8):e3001136. doi: 
            10.1371/journal.pbio.3001136.
  PUBMED    34424903
REFERENCE   2  (bases 1 to 8398)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS 
            Biol."; date: "2021-08-23"; volume: "19(8):e3001136. doi"; pages: " 
            10.1371/journal.pbio.3001136"
COMMENT     Sequence Label: pICSL86977OD:PITG_04266
FEATURES             Location/Qualifiers
     source          1..8398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(248..877)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    1186..1210
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1277..1984)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2397..2742)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2434..2454)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3775..4027)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4627..4809)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4663..4841)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5004..5028
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5164..5752)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5244..5263)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5923..6717)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6718..6822)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6884..7078
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7144..8217)
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
                     QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
                     KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
                     GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
                     ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
                     LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
                     LVMRYRNLIEGEASAGS"
ORIGIN
        1 catttgctca tttactctgg tagctgcgcg atgtattcag atagcagctc ggtaatggtc
       61 ttgccttggc gtaccgcgta catcttcagc ttggtgtgat cctccgccgg caactgaaag
      121 ttgacccgct tcatggctgg cgtgtctgcc aggctggcca acgttgcagc cttgctgctg
      181 cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt tgctcatttt ctctttacct
      241 cattaactca aatgagtttt gatttaattt cagcggccag cgcctggacc tcgcgggcag
      301 cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc
      361 tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc gtacccggcc agcgcctcgg
      421 caacctcacc gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta
      481 gccttccatc cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc
      541 atatgtcgta agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg
      601 acacagccaa gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg cgccggccga
      661 tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat
      721 cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg actaacagaa
      781 catcggcccc ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg
      841 acccgccttt ctggttaagt acagcgataa ccttcatgcg ttccccttgc gtatttgttt
      901 atttactcat cgcatcatat acgcagcgac cgcatgacgc aagctgtttt actcaaatac
      961 acatcacctt tttagacggc ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc
     1021 cgccagcttg gcatcagaca aaccggccag gatttcatgc agccgcacgg ctgactcccg
     1081 aggggaaccc tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc
     1141 ccttttaaat atccgattat tctaataaac gctcttttct cttaggttta cccgccaata
     1201 tatcctgtca aacactgata gtttaaacgg atcacttcgt gtccctcggt cacatgtgca
     1261 tcctctagta gcggtcctgc tgagcctcga catgttgtcg caaaattcgc cctggacccg
     1321 cccaacgatt tgtcgtcact gtcaaggttt gacctgcact tcatttgggg cccacataca
     1381 ccaaaaaaat gctgcataat tctcggggca gcaagtcggt tacccggccg ccgtgctgga
     1441 ccgggttgaa tggtgcccgt aactttcggt agagcggacg gccaatactc aacttcaagg
     1501 aatctcaccc atgcgcgccg gcggggaacc ggagttccct tcagtgagcg ttattagttc
     1561 gccgctcggt gtgtcgtaga tactagcccc tggggcactt ttgaaatttg aataagattt
     1621 atgtaatcag tcttttaggt ttgaccggtt ctgccgcttt ttttaaaatt ggatttgtaa
     1681 taataaaacg caattgtttg ttattgtggc gctctatcat agatgtcgct ataaacctat
     1741 tcagcacaat atattgtttt cattttaata ttgtacatat aagtagtagg gtacaatcag
     1801 taaattgaac ggagaatatt attcataaaa atacgatagt aacgggtgat atattcatta
     1861 gaatgaaccg aaaccggcgg taaggatctg agctacacat gctcaggttt tttacaacgt
     1921 gcacaacaga attgaaagca aatatcatgc gatcataggc ttctcgcata tctcattaaa
     1981 gcaggacaag ctcagtcaga gatcctgtcg atccatccaa gcaatcctcc cctagtaggc
     2041 ttaggagttg gagttggtgt aggagttgca gttgagttaa gagcaggagt aggtgtcaaa
     2101 gtctccttgt gcttcttctt tttcttcttt ctcttccttc tcttgaacaa tcccctctcg
     2161 tcgttgtcga aagaagagtt ctcgtcctcg tcctcgtact cattgtcctc gtccttccac
     2221 aacatgtcgt cgtcgtcgtg caaagtgtcg aacacgtatc cagttggagc cttagtagcc
     2281 tcgtaagtct tagactcaga gatcattgta tcgataattg taaatgtaat tgtaatgttg
     2341 tttgttgttt gttgttgttg gtaattgttg taaaaatgag ctcttatact cgagcgtgtc
     2401 ctctccaaat gaaatgaact tccttatata gaggaagggt cttgcgaagg atagtgggat
     2461 tgtgcgtcat cccttacgtc agtggagatg tcacatcaat ccacttgctt tgtagacgtg
     2521 gttggaacct cttctttttc cacgatgctc ctcgtgggtg ggggtccatc tttgggacca
     2581 ctgtcggcag agagatcttg aatgatagcc tttcctttat cgcaatgatg gcatttgtag
     2641 gagccacctt ccttttctac tgtcctttcg atgaagtgac agatagctgg gcaatggaat
     2701 ccgaggaggt ttcccgaaat tatcctttgt tgaaaagtct caatagccct ttgatcttct
     2761 gagactgtat ctttgacatt tttggagtag accagagtgt cgtgctccac catgttgacg
     2821 aagattttct tcttgtcatt gagtcgtaaa agactctgta tgaactgttc gccagtcttg
     2881 acggcgagtt ctgttagatc ctcgatttga atcttagact ccatgcatgg ccttagattc
     2941 agtaggaact acctttttag agactccaat ctctattact tgccttggtt tatgaagcaa
     3001 gccttgaatc gtccatactg gaatagtact tctgatcttg agaaatatgt ctttctctgt
     3061 gttcttgatg caattagtcc tgaatctttt gactgcatct ttaaccttct tgggaaggta
     3121 tttgatctcc tggagattgt tactcgggta gatcgtcttg atgagacttg ctgcgtaggc
     3181 ctctctaacc atctgtgggt cagcattctt tctgaaattg aagaggctaa ccttctcatt
     3241 atcagtggtg aacatagtgt cgtcaccttc accttcgaac ttccttccta gatcgtaaag
     3301 atagaggaaa tcgtccattg taatctccgg ggcaaaggag atctcttttg gggctggatc
     3361 actgctgggc cttttggttc ctagcgtgag ccagtgggct ttttgctttg gtgggcttgt
     3421 tagggcctta gcaaagctct tgggcttgag ttgagcttct cctttgggga tgaagttcaa
     3481 cctgtctgtt tgctgacttg ttgtgtacgc gtcagctgct gctcttgcct ctgtaatagt
     3541 ggcaaatttc ttgtgtgcaa ctccgggaac gccgtttgtt gccgcctttg tacaacccca
     3601 gtcatcgtat ataccggcat gtggaccgtt atacacaacg tagtagttga tatgagggtg
     3661 ttgaataccc gattctgctc tgagaggagc aactgtgctg ttaagctcag atttttgtgg
     3721 gattggaatt cctccaagct tgaattcttg cagcgctcca attcagcaag cgtcgatcta
     3781 gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt
     3841 ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata
     3901 acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga aattatatga
     3961 taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg
     4021 aacgatctgc ttgactctag atccagagtc ccgctcaaat ctcagtcaca ggaagaacag
     4081 gtctaggagg aacaggaaga gagaaatcaa gctgccagaa tcccacatca tgccaatttc
     4141 catgtttgaa accagcagct ctaagcattc ccctaggagc ataaccaaga gcctcatgca
     4201 tcctcacaga aggatcatta ggaagtccaa tcacagcaac cacagacttg aaaccctgag
     4261 cctccaaaga cttcaaaaga tgagtgtaaa gagtagatcc aagaccagtt ctttgatgcc
     4321 taggagacac gtaaacagta gactcagcag tccaatcgta agcattccta gccttccaag
     4381 gaccagcata agcaatacca gccacttcac catccacttc agcaacaagc caaggatacc
     4441 tctctctaag cctcacaaga tcatcagtcc actcttgtgg ttcttgaggc tcagtcctga
     4501 aattcacagt agaagtctca atgtaatgat tcacaatagt gcacacagca ggcatatcag
     4561 cctcagtagc ccttctaata tcagcaggcc ttctttcagg agacatgcga aacgatccag
     4621 atccggtgca gattatttgg attgagagtg aatatgagac tctaattgga taccgagggg
     4681 aatttatgga acgtcagtgg agcattttga caagaaatat ttgctagctg atagtgacct
     4741 taggcgactt ttgaacgcgc aataatggtt tctgacgtat gtgcttagct cattaaactc
     4801 cagaaacccg cggctgagtg gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa
     4861 acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt ctccgctccg
     4921 gatccgaatt cggcacagag tgttcaaccc cagtacatta aaaacgtccg caatgtgtta
     4981 ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc agccagccaa
     5041 cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag cccatcaacc
     5101 atgactactg tccggtccaa cagatccgag caggaaccgt aaaaaggccg cgttgctggc
     5161 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     5221 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     5281 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     5341 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     5401 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     5461 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     5521 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     5581 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     5641 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     5701 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     5761 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     5821 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     5881 taaatcaatc taaagtatat atgagtaaac ttggtctgac agctaaaaca attcatccag
     5941 taaaatataa tattttattt tctcccaatc aggcttgatc cccagtaagt caaaaaatag
     6001 ctcgacatac tgttcttccc cgatatcctc cctgagcgac cggacgcaga aggcaatgtc
     6061 ataccacttg tccgccctgc cgcttctccc aagatcaata aagccactta ctttgccatc
     6121 tttcacaaag atgttgctgt ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg
     6181 cttttccgtc tttaaaaaat catacagctc gcgcggatct ttaaatggag tgtcctcttc
     6241 ccagttttcg caatccacat cggccagatc gttattcagt aagtaatcca attcggctaa
     6301 gcggctgtct aagctattcg tatagggaca atccgatatg tcgatggagt gaaagagcct
     6361 gatgcactcc gcatacagct cgataatctt ttcagggctt tgttcatctt catactcttc
     6421 cgagcaaagg acgccatcgg cctcactcat gagcagattg ctccagccat catgccgttc
     6481 aaagtgcagg acctttggaa caggcagctt tccttccagc catagcatca tgtccttttc
     6541 ccgttccaca tcataggtgg tccctttata ccggctgtcc gtcattttta aatataggtt
     6601 ttcattttct cccaccagct tatatacctt agcaggagac attccttccg tatcttttac
     6661 gcagcggtat ttttcgatca gttttttcaa ttccggtgat attctcattt tagccatact
     6721 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     6781 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcactcgga gccttgatgt
     6841 gggcgctttt tgcagctgtt cggctgtgcg ctggccagac agttatgcac aggccaggcg
     6901 ggttttaaga gttttaataa gttttaaaga gttttaggcg gaaaaatcgc cttttttctc
     6961 ttttatatca gtcacttaca tgtgtgaccg gttcccaatg tacggctttg ggttcccaat
     7021 gtacgggttc cggttcccaa tgtacggctt tgggttccca atgtacgtgc tatccacagg
     7081 aaagaagctt tttcgacctt tttcccctgc tagggcaatt tgccctagca tctgctccgt
     7141 acattaggaa ccggcggatg cttcgccctc gatcaggttg cggtagcgca tgactaggat
     7201 cgggccagcc tgccccgcct cctccttcaa atcgtactcc ggcaggtcat ttgacccgat
     7261 cagcttgcgc acggtgaaac agaacttctt gaactctccg gcgctgccac tgcgttcgta
     7321 gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta
     7381 gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc
     7441 cgggttcttg ccttctgtga tctcgcggta catccaatca gctagctcga tctcgatgta
     7501 ctccggccgc ccggtttcgc tctttacgat cttgtagcgg ctaatcaagg cttcaccctc
     7561 ggataccgtc accaggcggc cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt
     7621 ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc tgctttccgc catcggctcg
     7681 ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc
     7741 cttcccttcc cggtatcggt tcatggattc ggttagatgg gaaaccgcca tcagtaccag
     7801 gtcgtaatcc cacacactgg ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc
     7861 tggaagctcg tagcggatca cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac
     7921 ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg tcatagagca tcggaacgaa
     7981 aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc gacgacgcac cggctgccgg
     8041 cggttgccgg gattctttgc ggattcgatc agcggccgct tgccacgatt caccggggcg
     8101 tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg gccttcaact tctccaccag
     8161 gtcatcaccc agcgccgcgc cgatttgtac cgggccggat ggtttgcgac cgctcacgcc
     8221 gattcctcgg gcttgggggt tccagtgcca ttgcagggcc ggcagacaac ccagccgctt
     8281 acgcctggcc aaccgcccgt tcctccacac atggggcatt ccacggcgtc ggtgcctggt
     8341 tgttcttgat tttccatgcc gcctccttta gccgctaaaa ttcatctact catttatt
//
