LOCUS       sequence-342206-        9070 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 9070)
  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 9070)
  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_10540
FEATURES             Location/Qualifiers
     source          1..9070
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(205..834)
                     /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    1143..1167
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1234..1941)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3026..3371)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3063..3083)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4404..4656)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5256..5438)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5292..5470)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5633..5657
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5793..6381)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5873..5892)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6552..7346)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7347..7451)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7513..7707
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7773..8846)
                     /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 gcagctcggt aatggtcttg ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct
       61 ccgccggcaa ctgaaagttg acccgcttca tggctggcgt gtctgccagg ctggccaacg
      121 ttgcagcctt gctgctgcgt gcgctcggac ggccggcact tagcgtgttt gtgcttttgc
      181 tcattttctc tttacctcat taactcaaat gagttttgat ttaatttcag cggccagcgc
      241 ctggacctcg cgggcagcgt cgccctcggg ttctgattca agaacggttg tgccggcggc
      301 ggcagtgcct gggtagctca cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta
      361 cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac
      421 gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa ccagctccac
      481 caggtcggcg gtggcccata tgtcgtaagg gcttggctgc accggaatca gcacgaagtc
      541 ggctgccttg atcgcggaca cagccaagtc cgccgcctgg ggcgctccgt cgatcactac
      601 gaagtcgcgc cggccgatgg ccttcacgtc gcggtcaatc gtcgggcggt cgatgccgac
      661 aacggttagc ggttgatctt cccgcacggc cgcccaatcg cgggcactgc cctggggatc
      721 ggaatcgact aacagaacat cggccccggc gagttgcagg gcgcgggcta gatgggttgc
      781 gatggtcgtc ttgcctgacc cgcctttctg gttaagtaca gcgataacct tcatgcgttc
      841 cccttgcgta tttgtttatt tactcatcgc atcatatacg cagcgaccgc atgacgcaag
      901 ctgttttact caaatacaca tcaccttttt agacggcggc gctcggtttc ttcagcggcc
      961 aagctggccg gccaggccgc cagcttggca tcagacaaac cggccaggat ttcatgcagc
     1021 cgcacggctg actcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg
     1081 gataaacctt ttcacgccct tttaaatatc cgattattct aataaacgct cttttctctt
     1141 aggtttaccc gccaatatat cctgtcaaac actgatagtt taaacggatc acttcgtgtc
     1201 cctcggtcac atgtgcatcc tctagtagcg gtcctgctga gcctcgacat gttgtcgcaa
     1261 aattcgccct ggacccgccc aacgatttgt cgtcactgtc aaggtttgac ctgcacttca
     1321 tttggggccc acatacacca aaaaaatgct gcataattct cggggcagca agtcggttac
     1381 ccggccgccg tgctggaccg ggttgaatgg tgcccgtaac tttcggtaga gcggacggcc
     1441 aatactcaac ttcaaggaat ctcacccatg cgcgccggcg gggaaccgga gttcccttca
     1501 gtgagcgtta ttagttcgcc gctcggtgtg tcgtagatac tagcccctgg ggcacttttg
     1561 aaatttgaat aagatttatg taatcagtct tttaggtttg accggttctg ccgctttttt
     1621 taaaattgga tttgtaataa taaaacgcaa ttgtttgtta ttgtggcgct ctatcataga
     1681 tgtcgctata aacctattca gcacaatata ttgttttcat tttaatattg tacatataag
     1741 tagtagggta caatcagtaa attgaacgga gaatattatt cataaaaata cgatagtaac
     1801 gggtgatata ttcattagaa tgaaccgaaa ccggcggtaa ggatctgagc tacacatgct
     1861 caggtttttt acaacgtgca caacagaatt gaaagcaaat atcatgcgat cataggcttc
     1921 tcgcatatct cattaaagca ggacaagctc aagccaaatt cttgttcaaa gccttagctt
     1981 ggatcaaaag gatcacagcc tggtattcct ttgaatcctt tggtgtagac ctagtaactt
     2041 taagctctct cttcaaagct gtcaaagtca atgattcgtc cataaactgt ccgaaatact
     2101 tttgagccaa tgcgttatcg tcaatcttag tcaagattcc ctcagtgtta gacctcacag
     2161 tagggttctt agccttagcc actccgttga tgtacatcct agattgaacc aatcctgaca
     2221 atgcgtcata cttagtagtg tctggagtca tacccttagt gatacccaat ttagactgca
     2281 atgtttgaat agaccacttc tcatcaacga attccttgaa gaacttctgt gccaaaggag
     2341 atccctccac ccttcccatg atgttctcct tcaatggaga accttttgtc tttgccaaca
     2401 ttgagttcca tgccctagcc tggataagca atgtcaaagc gtcgtattct tttgatgtct
     2461 taagagttcc tctagtgata ccaagtttgt tcttaagagt cacctgtgta agagtgtcgt
     2521 ccatgaactg tccgaagtac ttctgagcca atgggttttc gttgatctta gccaacatag
     2581 tctctcctgt attagttgta gttgggttct taaccttcat aagagtatcc acgtacatcc
     2641 tagtctgcaa caatgtagaa aatgcctcgt actcctttga gtctggagct gtgtccttag
     2701 taatcctcaa cttagactta agcacgtcaa cagtccactt ctcgtcgatg tactgctcga
     2761 agttcctctg tgccaactgg ttctttccca actttgacat caagttctgc tcaagtgaca
     2821 tagcgttctg aggctttgca gctgccactg cctcgtccac cttcttaagt gcaccagtgt
     2881 tctttgtaaa agaagccaac tttgaaagct gctttgtcaa gatttccatc ctcattgtat
     2941 cgataattgt aaatgtaatt gtaatgttgt ttgttgtttg ttgttgttgg taattgttgt
     3001 aaaaatgagc tcttatactc gagcgtgtcc tctccaaatg aaatgaactt ccttatatag
     3061 aggaagggtc ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca gtggagatgt
     3121 cacatcaatc cacttgcttt gtagacgtgg ttggaacctc ttctttttcc acgatgctcc
     3181 tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga gagatcttga atgatagcct
     3241 ttcctttatc gcaatgatgg catttgtagg agccaccttc cttttctact gtcctttcga
     3301 tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgaaatt atcctttgtt
     3361 gaaaagtctc aatagccctt tgatcttctg agactgtatc tttgacattt ttggagtaga
     3421 ccagagtgtc gtgctccacc atgttgacga agattttctt cttgtcattg agtcgtaaaa
     3481 gactctgtat gaactgttcg ccagtcttga cggcgagttc tgttagatcc tcgatttgaa
     3541 tcttagactc catgcatggc cttagattca gtaggaacta cctttttaga gactccaatc
     3601 tctattactt gccttggttt atgaagcaag ccttgaatcg tccatactgg aatagtactt
     3661 ctgatcttga gaaatatgtc tttctctgtg ttcttgatgc aattagtcct gaatcttttg
     3721 actgcatctt taaccttctt gggaaggtat ttgatctcct ggagattgtt actcgggtag
     3781 atcgtcttga tgagacttgc tgcgtaggcc tctctaacca tctgtgggtc agcattcttt
     3841 ctgaaattga agaggctaac cttctcatta tcagtggtga acatagtgtc gtcaccttca
     3901 ccttcgaact tccttcctag atcgtaaaga tagaggaaat cgtccattgt aatctccggg
     3961 gcaaaggaga tctcttttgg ggctggatca ctgctgggcc ttttggttcc tagcgtgagc
     4021 cagtgggctt tttgctttgg tgggcttgtt agggccttag caaagctctt gggcttgagt
     4081 tgagcttctc ctttggggat gaagttcaac ctgtctgttt gctgacttgt tgtgtacgcg
     4141 tcagctgctg ctcttgcctc tgtaatagtg gcaaatttct tgtgtgcaac tccgggaacg
     4201 ccgtttgttg ccgcctttgt acaaccccag tcatcgtata taccggcatg tggaccgtta
     4261 tacacaacgt agtagttgat atgagggtgt tgaatacccg attctgctct gagaggagca
     4321 actgtgctgt taagctcaga tttttgtggg attggaattc ctccaagctt gaattcttgc
     4381 agcgctccaa ttcagcaagc gtcgatctag taacatagat gacaccgcgc gcgataattt
     4441 atcctagttt gcgcgctata ttttgttttc tatcgcgtat taaatgtata attgcgggac
     4501 tctaatcata aaaacccatc tcataaataa cgtcatgcat tacatgttaa ttattacatg
     4561 cttaacgtaa ttcaacagaa attatatgat aatcatcgca agaccggcaa caggattcaa
     4621 tcttaagaaa ctttattgcc aaatgtttga acgatctgct tgactctaga tccagagtcc
     4681 cgctcaaatc tcagtcacag gaagaacagg tctaggagga acaggaagag agaaatcaag
     4741 ctgccagaat cccacatcat gccaatttcc atgtttgaaa ccagcagctc taagcattcc
     4801 cctaggagca taaccaagag cctcatgcat cctcacagaa ggatcattag gaagtccaat
     4861 cacagcaacc acagacttga aaccctgagc ctccaaagac ttcaaaagat gagtgtaaag
     4921 agtagatcca agaccagttc tttgatgcct aggagacacg taaacagtag actcagcagt
     4981 ccaatcgtaa gcattcctag ccttccaagg accagcataa gcaataccag ccacttcacc
     5041 atccacttca gcaacaagcc aaggatacct ctctctaagc ctcacaagat catcagtcca
     5101 ctcttgtggt tcttgaggct cagtcctgaa attcacagta gaagtctcaa tgtaatgatt
     5161 cacaatagtg cacacagcag gcatatcagc ctcagtagcc cttctaatat cagcaggcct
     5221 tctttcagga gacatgcgaa acgatccaga tccggtgcag attatttgga ttgagagtga
     5281 atatgagact ctaattggat accgagggga atttatggaa cgtcagtgga gcattttgac
     5341 aagaaatatt tgctagctga tagtgacctt aggcgacttt tgaacgcgca ataatggttt
     5401 ctgacgtatg tgcttagctc attaaactcc agaaacccgc ggctgagtgg ctccttcaac
     5461 gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca
     5521 taacgtgact cccttaattc tccgctccgg atccgaattc ggcacagagt gttcaacccc
     5581 agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac
     5641 cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat
     5701 caccactcga tacaggcagc ccatcaacca tgactactgt ccggtccaac agatccgagc
     5761 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5821 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5881 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5941 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6001 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6061 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6121 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6181 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6241 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6301 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     6361 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     6421 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     6481 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     6541 tggtctgaca gctaaaacaa ttcatccagt aaaatataat attttatttt ctcccaatca
     6601 ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc gatatcctcc
     6661 ctgagcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc gcttctccca
     6721 agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg
     6781 ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg
     6841 cgcggatctt taaatggagt gtcctcttcc cagttttcgc aatccacatc ggccagatcg
     6901 ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt atagggacaa
     6961 tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc gataatcttt
     7021 tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc ctcactcatg
     7081 agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac aggcagcttt
     7141 ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt ccctttatac
     7201 cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt atatacctta
     7261 gcaggagaca ttccttccgt atcttttacg cagcggtatt tttcgatcag ttttttcaat
     7321 tccggtgata ttctcatttt agccatactc ttcctttttc aatattattg aagcatttat
     7381 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7441 ggggttccgc gcactcggag ccttgatgtg ggcgcttttt gcagctgttc ggctgtgcgc
     7501 tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag
     7561 ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg
     7621 ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt
     7681 gggttcccaa tgtacgtgct atccacagga aagaagcttt ttcgaccttt ttcccctgct
     7741 agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg
     7801 atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa
     7861 tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg
     7921 aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc
     7981 ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa
     8041 aagtaatcgg ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat ctcgcggtac
     8101 atccaatcag ctagctcgat ctcgatgtac tccggccgcc cggtttcgct ctttacgatc
     8161 ttgtagcggc taatcaaggc ttcaccctcg gataccgtca ccaggcggcc gttcttggcc
     8221 ttcttcgtac gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt ttctaccagg
     8281 tcgtctttct gctttccgcc atcggctcgc cggcagaact tgagtacgtc cgcaacgtgt
     8341 ggacggaaca cgcggccggg cttgtctccc ttcccttccc ggtatcggtt catggattcg
     8401 gttagatggg aaaccgccat cagtaccagg tcgtaatccc acacactggc catgccggcc
     8461 ggccctgcgg aaacctctac gtgcccgtct ggaagctcgt agcggatcac ctcgccagct
     8521 cgtcggtcac gcttcgacag acggaaaacg gccacgtcca tgatgctgcg actatcgcgg
     8581 gtgcccacgt catagagcat cggaacgaaa aaatctggtt gctcgtcgcc cttgggcggc
     8641 ttcctaatcg acgacgcacc ggctgccggc ggttgccggg attctttgcg gattcgatca
     8701 gcggccgctt gccacgattc accggggcgt gcttctgcct cgatgcgttg ccgctgggcg
     8761 gcctgcgcgg ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc gatttgtacc
     8821 gggccggatg gtttgcgacc gctcacgccg attcctcggg cttgggggtt ccagtgccat
     8881 tgcagggccg gcagacaacc cagccgctta cgcctggcca accgcccgtt cctccacaca
     8941 tggggcattc cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg cctcctttag
     9001 ccgctaaaat tcatctactc atttattcat ttgctcattt actctggtag ctgcgcgatg
     9061 tattcagata
//
