LOCUS       sequence-342346-        9484 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 9484)
  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 9484)
  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_15123a
FEATURES             Location/Qualifiers
     source          1..9484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(243..872)
                     /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    1181..1205
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1272..1979)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3478..3823)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3515..3535)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4856..5108)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5708..5890)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5744..5922)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6085..6109
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6245..6833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6325..6344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7004..7798)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7799..7903)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7965..8159
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8225..9298)
                     /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 gctcatttac tctggtagct gcgcgatgta ttcagatagc agctcggtaa tggtcttgcc
       61 ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc gccggcaact gaaagttgac
      121 ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt gcagccttgc tgctgcgtgc
      181 gctcggacgg ccggcactta gcgtgtttgt gcttttgctc attttctctt tacctcatta
      241 actcaaatga gttttgattt aatttcagcg gccagcgcct ggacctcgcg ggcagcgtcg
      301 ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg gtagctcacg
      361 cgctgcgtga tacgggactc aagaatgggc agctcgtacc cggccagcgc ctcggcaacc
      421 tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc ttgtagcctt
      481 ccatccgtga cctcaatgcg ctgcttaacc agctccacca ggtcggcggt ggcccatatg
      541 tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg ctgccttgat cgcggacaca
      601 gccaagtccg ccgcctgggg cgctccgtcg atcactacga agtcgcgccg gccgatggcc
      661 ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg ttgatcttcc
      721 cgcacggccg cccaatcgcg ggcactgccc tggggatcgg aatcgactaa cagaacatcg
      781 gccccggcga gttgcagggc gcgggctaga tgggttgcga tggtcgtctt gcctgacccg
      841 cctttctggt taagtacagc gataaccttc atgcgttccc cttgcgtatt tgtttattta
      901 ctcatcgcat catatacgca gcgaccgcat gacgcaagct gttttactca aatacacatc
      961 acctttttag acggcggcgc tcggtttctt cagcggccaa gctggccggc caggccgcca
     1021 gcttggcatc agacaaaccg gccaggattt catgcagccg cacggctgac tcccgagggg
     1081 aaccctgtgg ttggcatgca catacaaatg gacgaacgga taaacctttt cacgcccttt
     1141 taaatatccg attattctaa taaacgctct tttctcttag gtttacccgc caatatatcc
     1201 tgtcaaacac tgatagttta aacggatcac ttcgtgtccc tcggtcacat gtgcatcctc
     1261 tagtagcggt cctgctgagc ctcgacatgt tgtcgcaaaa ttcgccctgg acccgcccaa
     1321 cgatttgtcg tcactgtcaa ggtttgacct gcacttcatt tggggcccac atacaccaaa
     1381 aaaatgctgc ataattctcg gggcagcaag tcggttaccc ggccgccgtg ctggaccggg
     1441 ttgaatggtg cccgtaactt tcggtagagc ggacggccaa tactcaactt caaggaatct
     1501 cacccatgcg cgccggcggg gaaccggagt tcccttcagt gagcgttatt agttcgccgc
     1561 tcggtgtgtc gtagatacta gcccctgggg cacttttgaa atttgaataa gatttatgta
     1621 atcagtcttt taggtttgac cggttctgcc gcttttttta aaattggatt tgtaataata
     1681 aaacgcaatt gtttgttatt gtggcgctct atcatagatg tcgctataaa cctattcagc
     1741 acaatatatt gttttcattt taatattgta catataagta gtagggtaca atcagtaaat
     1801 tgaacggaga atattattca taaaaatacg atagtaacgg gtgatatatt cattagaatg
     1861 aaccgaaacc ggcggtaagg atctgagcta cacatgctca ggttttttac aacgtgcaca
     1921 acagaattga aagcaaatat catgcgatca taggcttctc gcatatctca ttaaagcagg
     1981 acaagctcac ctccttgggt tgattgatgt gtcgatgtga ggaggtgcca ttttcttctt
     2041 gtaatagtcc ttgtatctag cagcagtagc cctctccaag ttagatgctg tgttctctga
     2101 tgatctgaaa attgtaccat aaatatcaga aggctctgtc tccctcttga accacctacc
     2161 gaacaacatc ctttggtact cagtagcgat tgagatttcg tccaagtcag cgtgtccgtt
     2221 cctaatcaag tgctcccttg ccctagttgt ctgaattgcc aactcgtcct cacctccgaa
     2281 acccttagac actgccctca acataccctc gttgtacaaa aggttcttgt tgttcaaagt
     2341 agccaaataa ccagaaagag tgtcgaactt tggattagta ggaggaagca aatcgttaaa
     2401 gtgaatgtcc aaaagtctga acacagagtt tgcagagatt ccaccctctt tccacttcaa
     2461 gaattgttgc ttttgcaatg tgataaccaa gttcattgta gctgcgtcgt caggcttagc
     2521 gaatgtaagc acagttgcca ctgctttatt tccgtactga gccacaaatg agtcgatcaa
     2581 agatgccttt gaatttgggt tttttgcgtt atacattgag atgaaagctc cgaaagtgtg
     2641 cacgttcctg tcaagcaatg cgtccaatcc cctgttcaac ctcaactttt tgtagatatg
     2701 tttaggtcta agaagctcca ccctcaccca cttgttgata agagcgttct gcaaagctga
     2761 agcaagctcc cttgtcttaa cgtcctccct tgctctagca aggattgttg caagctgctc
     2821 ctctccacca aatcctttag tcaaagtttt caaaagtgag tctttagtag actttgttga
     2881 aataaccaat ttaatgtagt cgtcaagcac ttccatctta ggagaagcca atgccaaata
     2941 tccatcagac ctcaatttaa gcaaagtaaa aacgtcatca actgatttcc tcttctgcaa
     3001 ccaagaaagc aactgagcgt gcctaagcct ctggatagtg ttcatctcac caccagccct
     3061 ctgtgcagtc atcaatgcgt ttgccacagc gtcgtcaccg tagtgagcca tgaacatacc
     3121 gatcactgaa gccttgttgt tgttgttctg agtattcaac tctttaacat aattttccaa
     3181 tgccttaagg ttaggtgaag caagagcgtc gtccattccg tttgtcaact tgaacctcct
     3241 caagataggc tcagctggag ggtccttaga gatccatgat ccaaggttga tcttagacac
     3301 tccagcctta gcaaggtctg taagcttctc caatcctgct gagatcaccc tgttctcacc
     3361 attctcagtg ttctgtgtag acaccattgt atcgataatt gtaaatgtaa ttgtaatgtt
     3421 gtttgttgtt tgttgttgtt ggtaattgtt gtaaaaatga gctcttatac tcgagcgtgt
     3481 cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag gatagtggga
     3541 ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct ttgtagacgt
     3601 ggttggaacc tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc
     3661 actgtcggca gagagatctt gaatgatagc ctttccttta tcgcaatgat ggcatttgta
     3721 ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg ggcaatggaa
     3781 tccgaggagg tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc tttgatcttc
     3841 tgagactgta tctttgacat ttttggagta gaccagagtg tcgtgctcca ccatgttgac
     3901 gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt cgccagtctt
     3961 gacggcgagt tctgttagat cctcgatttg aatcttagac tccatgcatg gccttagatt
     4021 cagtaggaac taccttttta gagactccaa tctctattac ttgccttggt ttatgaagca
     4081 agccttgaat cgtccatact ggaatagtac ttctgatctt gagaaatatg tctttctctg
     4141 tgttcttgat gcaattagtc ctgaatcttt tgactgcatc tttaaccttc ttgggaaggt
     4201 atttgatctc ctggagattg ttactcgggt agatcgtctt gatgagactt gctgcgtagg
     4261 cctctctaac catctgtggg tcagcattct ttctgaaatt gaagaggcta accttctcat
     4321 tatcagtggt gaacatagtg tcgtcacctt caccttcgaa cttccttcct agatcgtaaa
     4381 gatagaggaa atcgtccatt gtaatctccg gggcaaagga gatctctttt ggggctggat
     4441 cactgctggg ccttttggtt cctagcgtga gccagtgggc tttttgcttt ggtgggcttg
     4501 ttagggcctt agcaaagctc ttgggcttga gttgagcttc tcctttgggg atgaagttca
     4561 acctgtctgt ttgctgactt gttgtgtacg cgtcagctgc tgctcttgcc tctgtaatag
     4621 tggcaaattt cttgtgtgca actccgggaa cgccgtttgt tgccgccttt gtacaacccc
     4681 agtcatcgta tataccggca tgtggaccgt tatacacaac gtagtagttg atatgagggt
     4741 gttgaatacc cgattctgct ctgagaggag caactgtgct gttaagctca gatttttgtg
     4801 ggattggaat tcctccaagc ttgaattctt gcagcgctcc aattcagcaa gcgtcgatct
     4861 agtaacatag atgacaccgc gcgcgataat ttatcctagt ttgcgcgcta tattttgttt
     4921 tctatcgcgt attaaatgta taattgcggg actctaatca taaaaaccca tctcataaat
     4981 aacgtcatgc attacatgtt aattattaca tgcttaacgt aattcaacag aaattatatg
     5041 ataatcatcg caagaccggc aacaggattc aatcttaaga aactttattg ccaaatgttt
     5101 gaacgatctg cttgactcta gatccagagt cccgctcaaa tctcagtcac aggaagaaca
     5161 ggtctaggag gaacaggaag agagaaatca agctgccaga atcccacatc atgccaattt
     5221 ccatgtttga aaccagcagc tctaagcatt cccctaggag cataaccaag agcctcatgc
     5281 atcctcacag aaggatcatt aggaagtcca atcacagcaa ccacagactt gaaaccctga
     5341 gcctccaaag acttcaaaag atgagtgtaa agagtagatc caagaccagt tctttgatgc
     5401 ctaggagaca cgtaaacagt agactcagca gtccaatcgt aagcattcct agccttccaa
     5461 ggaccagcat aagcaatacc agccacttca ccatccactt cagcaacaag ccaaggatac
     5521 ctctctctaa gcctcacaag atcatcagtc cactcttgtg gttcttgagg ctcagtcctg
     5581 aaattcacag tagaagtctc aatgtaatga ttcacaatag tgcacacagc aggcatatca
     5641 gcctcagtag cccttctaat atcagcaggc cttctttcag gagacatgcg aaacgatcca
     5701 gatccggtgc agattatttg gattgagagt gaatatgaga ctctaattgg ataccgaggg
     5761 gaatttatgg aacgtcagtg gagcattttg acaagaaata tttgctagct gatagtgacc
     5821 ttaggcgact tttgaacgcg caataatggt ttctgacgta tgtgcttagc tcattaaact
     5881 ccagaaaccc gcggctgagt ggctccttca acgttgcggt tctgtcagtt ccaaacgtaa
     5941 aacggcttgt cccgcgtcat cggcgggggt cataacgtga ctcccttaat tctccgctcc
     6001 ggatccgaat tcggcacaga gtgttcaacc ccagtacatt aaaaacgtcc gcaatgtgtt
     6061 attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac cagccagcca
     6121 acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca gcccatcaac
     6181 catgactact gtccggtcca acagatccga gcaggaaccg taaaaaggcc gcgttgctgg
     6241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     6301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     6361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     6421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     6481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     6541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     6601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     6661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     6721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6841 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6901 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6961 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagctaaaac aattcatcca
     7021 gtaaaatata atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata
     7081 gctcgacata ctgttcttcc ccgatatcct ccctgagcga ccggacgcag aaggcaatgt
     7141 cataccactt gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat
     7201 ctttcacaaa gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg
     7261 gcttttccgt ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcctctt
     7321 cccagttttc gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta
     7381 agcggctgtc taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc
     7441 tgatgcactc cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt
     7501 ccgagcaaag gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt
     7561 caaagtgcag gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt
     7621 cccgttccac atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt
     7681 tttcattttc tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta
     7741 cgcagcggta tttttcgatc agttttttca attccggtga tattctcatt ttagccatac
     7801 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7861 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcactcgg agccttgatg
     7921 tgggcgcttt ttgcagctgt tcggctgtgc gctggccaga cagttatgca caggccaggc
     7981 gggttttaag agttttaata agttttaaag agttttaggc ggaaaaatcg ccttttttct
     8041 cttttatatc agtcacttac atgtgtgacc ggttcccaat gtacggcttt gggttcccaa
     8101 tgtacgggtt ccggttccca atgtacggct ttgggttccc aatgtacgtg ctatccacag
     8161 gaaagaagct ttttcgacct ttttcccctg ctagggcaat ttgccctagc atctgctccg
     8221 tacattagga accggcggat gcttcgccct cgatcaggtt gcggtagcgc atgactagga
     8281 tcgggccagc ctgccccgcc tcctccttca aatcgtactc cggcaggtca tttgacccga
     8341 tcagcttgcg cacggtgaaa cagaacttct tgaactctcc ggcgctgcca ctgcgttcgt
     8401 agatcgtctt gaacaaccat ctggcttctg ccttgcctgc ggcgcggcgt gccaggcggt
     8461 agagaaaacg gccgatgccg ggatcgatca aaaagtaatc ggggtgaacc gtcagcacgt
     8521 ccgggttctt gccttctgtg atctcgcggt acatccaatc agctagctcg atctcgatgt
     8581 actccggccg cccggtttcg ctctttacga tcttgtagcg gctaatcaag gcttcaccct
     8641 cggataccgt caccaggcgg ccgttcttgg ccttcttcgt acgctgcatg gcaacgtgcg
     8701 tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt ctgctttccg ccatcggctc
     8761 gccggcagaa cttgagtacg tccgcaacgt gtggacggaa cacgcggccg ggcttgtctc
     8821 ccttcccttc ccggtatcgg ttcatggatt cggttagatg ggaaaccgcc atcagtacca
     8881 ggtcgtaatc ccacacactg gccatgccgg ccggccctgc ggaaacctct acgtgcccgt
     8941 ctggaagctc gtagcggatc acctcgccag ctcgtcggtc acgcttcgac agacggaaaa
     9001 cggccacgtc catgatgctg cgactatcgc gggtgcccac gtcatagagc atcggaacga
     9061 aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat cgacgacgca ccggctgccg
     9121 gcggttgccg ggattctttg cggattcgat cagcggccgc ttgccacgat tcaccggggc
     9181 gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc ggccttcaac ttctccacca
     9241 ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga tggtttgcga ccgctcacgc
     9301 cgattcctcg ggcttggggg ttccagtgcc attgcagggc cggcagacaa cccagccgct
     9361 tacgcctggc caaccgcccg ttcctccaca catggggcat tccacggcgt cggtgcctgg
     9421 ttgttcttga ttttccatgc cgcctccttt agccgctaaa attcatctac tcatttattc
     9481 attt
//
