LOCUS       sequence-342342-        9724 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 9724)
  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 9724)
  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_14884
FEATURES             Location/Qualifiers
     source          1..9724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(273..902)
                     /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    1211..1235
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1302..2009)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3748..4093)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3785..3805)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(5126..5378)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5978..6160)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(6014..6192)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6355..6379
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6515..7103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6595..6614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7274..8068)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(8069..8173)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8235..8429
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8495..9568)
                     /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 gctaaaattc atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgta
       61 ttcagatagc agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt
      121 gtgatcctcc gccggcaact gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct
      181 ggccaacgtt gcagccttgc tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt
      241 gcttttgctc attttctctt tacctcatta actcaaatga gttttgattt aatttcagcg
      301 gccagcgcct ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg
      361 ccggcggcgg cagtgcctgg gtagctcacg cgctgcgtga tacgggactc aagaatgggc
      421 agctcgtacc cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc
      481 cgcgacacga caaaggccgc ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc
      541 agctccacca ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac cggaatcagc
      601 acgaagtcgg ctgccttgat cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg
      661 atcactacga agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg
      721 atgccgacaa cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc
      781 tggggatcgg aatcgactaa cagaacatcg gccccggcga gttgcagggc gcgggctaga
      841 tgggttgcga tggtcgtctt gcctgacccg cctttctggt taagtacagc gataaccttc
      901 atgcgttccc cttgcgtatt tgtttattta ctcatcgcat catatacgca gcgaccgcat
      961 gacgcaagct gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt
     1021 cagcggccaa gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt
     1081 catgcagccg cacggctgac tcccgagggg aaccctgtgg ttggcatgca catacaaatg
     1141 gacgaacgga taaacctttt cacgcccttt taaatatccg attattctaa taaacgctct
     1201 tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta aacggatcac
     1261 ttcgtgtccc tcggtcacat gtgcatcctc tagtagcggt cctgctgagc ctcgacatgt
     1321 tgtcgcaaaa ttcgccctgg acccgcccaa cgatttgtcg tcactgtcaa ggtttgacct
     1381 gcacttcatt tggggcccac atacaccaaa aaaatgctgc ataattctcg gggcagcaag
     1441 tcggttaccc ggccgccgtg ctggaccggg ttgaatggtg cccgtaactt tcggtagagc
     1501 ggacggccaa tactcaactt caaggaatct cacccatgcg cgccggcggg gaaccggagt
     1561 tcccttcagt gagcgttatt agttcgccgc tcggtgtgtc gtagatacta gcccctgggg
     1621 cacttttgaa atttgaataa gatttatgta atcagtcttt taggtttgac cggttctgcc
     1681 gcttttttta aaattggatt tgtaataata aaacgcaatt gtttgttatt gtggcgctct
     1741 atcatagatg tcgctataaa cctattcagc acaatatatt gttttcattt taatattgta
     1801 catataagta gtagggtaca atcagtaaat tgaacggaga atattattca taaaaatacg
     1861 atagtaacgg gtgatatatt cattagaatg aaccgaaacc ggcggtaagg atctgagcta
     1921 cacatgctca ggttttttac aacgtgcaca acagaattga aagcaaatat catgcgatca
     1981 taggcttctc gcatatctca ttaaagcagg acaagctcag tacaatggag atcctccatt
     2041 agccttgtag aaatcgtaga atccaagcac aactccgttg ttcctgatgt cccttggttg
     2101 ctcagcaacc aatgcagcaa ccctacctgg gtccaacctt ttttggttca tccactggtc
     2161 gaactgctgg ttcctcaatg aaaacaatct ttccactgag tgtcccttca tctctgcctg
     2221 gatctttgcg tgtgacaaca tcctagccaa gtccacgtag tcgaaccttt tctccaactc
     2281 tgtgaaaaga gcgaacttat cagggtgctt ctcatacttg ttaagcctct caacgtaaag
     2341 aatccatgcg tcgaacatag gagcttcgaa caagtcgtca cctgtcttgt caagttggaa
     2401 gaacttgaaa atgtcatctg ctgacttacc aggagatctc cacatctctg cagcaagctt
     2461 tgaagcaagt gcccttgtca acactgaagt ctttgcctgt gcgatcatct tagcaaggat
     2521 ctcgtcagag tagtgtttct tcataacaga gaaaaccaat gcgtctgtat ttgtgtagtc
     2581 cacaagctta acataagaaa cccaagtgtt gaacacagaa gattccaaaa gttcctcacc
     2641 ctccttagac aatccaaggt gcctgaagat atcgtcagca gtgtcacccc ttccgtgcca
     2701 cttagacaac tgaagcttct gcaaagcagt aacgttgatt ttcacatttt tgtccctaga
     2761 tgcagccacc aacacccttg ccaattcagc gtcagagaac ctgttctgca attcaaggaa
     2821 caaaaccttg tttgggtcct tgttcctctg cctaatataa gaaagccatg taggaagagc
     2881 agctgaccta aggaatgact ctcccttgtc agcgttcaac ctcaaaagct ttgccacgtc
     2941 accagtagtc tttccgtccc taagccaagc tgtcaaaagt gcgttctcca agttcctaac
     3001 cacagatgaa gtaagtgagt ctcccttagc tgcgataagc atgtgagcca atgcctcctc
     3061 gtcgttagtc ttagtaagtt taagcaacaa caagttataa gggtcctcgt tatccaattt
     3121 tttaacgaat gaaacccatg tattcaacaa tggattcctc aaaagacctt ctttgtcagc
     3181 gtcaagtttc aacaatttgt agatgtcctc ctctgtcttt ctgtttgtca cccagttagc
     3241 gatctgcatt gactcaattt tcttagcagt cctgattgct gctgcagttg gctgtgcctc
     3301 agacacaagc tttgccaaga tgtcatctcc gtggtgcctc acaagtgttg agaagatagc
     3361 tgcctgaccc ttctctggat tcttcttata agatttcatc actgatgcag cccacttctg
     3421 gtaaggagta gaagcgaaca actttgactc cacctcccac actcccaaag atgcaaacaa
     3481 cttatctgtt gctctttggt tatttgatgt caaaatttcc attgcccttg ttgacaattt
     3541 tttagttggc tgagagttga tcaacttctg caaaacagtt gctgcagaca acttaacacc
     3601 gatagcccta gcctctggag aaccgtcctc ttgctcagct gtccatttag actccattgt
     3661 atcgataatt gtaaatgtaa ttgtaatgtt gtttgttgtt tgttgttgtt ggtaattgtt
     3721 gtaaaaatga gctcttatac tcgagcgtgt cctctccaaa tgaaatgaac ttccttatat
     3781 agaggaaggg tcttgcgaag gatagtggga ttgtgcgtca tcccttacgt cagtggagat
     3841 gtcacatcaa tccacttgct ttgtagacgt ggttggaacc tcttcttttt ccacgatgct
     3901 cctcgtgggt gggggtccat ctttgggacc actgtcggca gagagatctt gaatgatagc
     3961 ctttccttta tcgcaatgat ggcatttgta ggagccacct tccttttcta ctgtcctttc
     4021 gatgaagtga cagatagctg ggcaatggaa tccgaggagg tttcccgaaa ttatcctttg
     4081 ttgaaaagtc tcaatagccc tttgatcttc tgagactgta tctttgacat ttttggagta
     4141 gaccagagtg tcgtgctcca ccatgttgac gaagattttc ttcttgtcat tgagtcgtaa
     4201 aagactctgt atgaactgtt cgccagtctt gacggcgagt tctgttagat cctcgatttg
     4261 aatcttagac tccatgcatg gccttagatt cagtaggaac taccttttta gagactccaa
     4321 tctctattac ttgccttggt ttatgaagca agccttgaat cgtccatact ggaatagtac
     4381 ttctgatctt gagaaatatg tctttctctg tgttcttgat gcaattagtc ctgaatcttt
     4441 tgactgcatc tttaaccttc ttgggaaggt atttgatctc ctggagattg ttactcgggt
     4501 agatcgtctt gatgagactt gctgcgtagg cctctctaac catctgtggg tcagcattct
     4561 ttctgaaatt gaagaggcta accttctcat tatcagtggt gaacatagtg tcgtcacctt
     4621 caccttcgaa cttccttcct agatcgtaaa gatagaggaa atcgtccatt gtaatctccg
     4681 gggcaaagga gatctctttt ggggctggat cactgctggg ccttttggtt cctagcgtga
     4741 gccagtgggc tttttgcttt ggtgggcttg ttagggcctt agcaaagctc ttgggcttga
     4801 gttgagcttc tcctttgggg atgaagttca acctgtctgt ttgctgactt gttgtgtacg
     4861 cgtcagctgc tgctcttgcc tctgtaatag tggcaaattt cttgtgtgca actccgggaa
     4921 cgccgtttgt tgccgccttt gtacaacccc agtcatcgta tataccggca tgtggaccgt
     4981 tatacacaac gtagtagttg atatgagggt gttgaatacc cgattctgct ctgagaggag
     5041 caactgtgct gttaagctca gatttttgtg ggattggaat tcctccaagc ttgaattctt
     5101 gcagcgctcc aattcagcaa gcgtcgatct agtaacatag atgacaccgc gcgcgataat
     5161 ttatcctagt ttgcgcgcta tattttgttt tctatcgcgt attaaatgta taattgcggg
     5221 actctaatca taaaaaccca tctcataaat aacgtcatgc attacatgtt aattattaca
     5281 tgcttaacgt aattcaacag aaattatatg ataatcatcg caagaccggc aacaggattc
     5341 aatcttaaga aactttattg ccaaatgttt gaacgatctg cttgactcta gatccagagt
     5401 cccgctcaaa tctcagtcac aggaagaaca ggtctaggag gaacaggaag agagaaatca
     5461 agctgccaga atcccacatc atgccaattt ccatgtttga aaccagcagc tctaagcatt
     5521 cccctaggag cataaccaag agcctcatgc atcctcacag aaggatcatt aggaagtcca
     5581 atcacagcaa ccacagactt gaaaccctga gcctccaaag acttcaaaag atgagtgtaa
     5641 agagtagatc caagaccagt tctttgatgc ctaggagaca cgtaaacagt agactcagca
     5701 gtccaatcgt aagcattcct agccttccaa ggaccagcat aagcaatacc agccacttca
     5761 ccatccactt cagcaacaag ccaaggatac ctctctctaa gcctcacaag atcatcagtc
     5821 cactcttgtg gttcttgagg ctcagtcctg aaattcacag tagaagtctc aatgtaatga
     5881 ttcacaatag tgcacacagc aggcatatca gcctcagtag cccttctaat atcagcaggc
     5941 cttctttcag gagacatgcg aaacgatcca gatccggtgc agattatttg gattgagagt
     6001 gaatatgaga ctctaattgg ataccgaggg gaatttatgg aacgtcagtg gagcattttg
     6061 acaagaaata tttgctagct gatagtgacc ttaggcgact tttgaacgcg caataatggt
     6121 ttctgacgta tgtgcttagc tcattaaact ccagaaaccc gcggctgagt ggctccttca
     6181 acgttgcggt tctgtcagtt ccaaacgtaa aacggcttgt cccgcgtcat cggcgggggt
     6241 cataacgtga ctcccttaat tctccgctcc ggatccgaat tcggcacaga gtgttcaacc
     6301 ccagtacatt aaaaacgtcc gcaatgtgtt attaagttgt ctaagcgtca atttgtttac
     6361 accacaatat atcctgccac cagccagcca acagctcccc gaccggcagc tcggcacaaa
     6421 atcaccactc gatacaggca gcccatcaac catgactact gtccggtcca acagatccga
     6481 gcaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     6541 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     6601 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     6661 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     6721 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6781 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6841 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6901 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6961 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     7021 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     7081 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     7141 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     7201 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     7261 cttggtctga cagctaaaac aattcatcca gtaaaatata atattttatt ttctcccaat
     7321 caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct
     7381 ccctgagcga ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc
     7441 caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt
     7501 cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct
     7561 cgcgcggatc tttaaatgga gtgtcctctt cccagttttc gcaatccaca tcggccagat
     7621 cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac
     7681 aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct
     7741 tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca
     7801 tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct
     7861 ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat
     7921 accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct
     7981 tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca
     8041 attccggtga tattctcatt ttagccatac tcttcctttt tcaatattat tgaagcattt
     8101 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8161 taggggttcc gcgcactcgg agccttgatg tgggcgcttt ttgcagctgt tcggctgtgc
     8221 gctggccaga cagttatgca caggccaggc gggttttaag agttttaata agttttaaag
     8281 agttttaggc ggaaaaatcg ccttttttct cttttatatc agtcacttac atgtgtgacc
     8341 ggttcccaat gtacggcttt gggttcccaa tgtacgggtt ccggttccca atgtacggct
     8401 ttgggttccc aatgtacgtg ctatccacag gaaagaagct ttttcgacct ttttcccctg
     8461 ctagggcaat ttgccctagc atctgctccg tacattagga accggcggat gcttcgccct
     8521 cgatcaggtt gcggtagcgc atgactagga tcgggccagc ctgccccgcc tcctccttca
     8581 aatcgtactc cggcaggtca tttgacccga tcagcttgcg cacggtgaaa cagaacttct
     8641 tgaactctcc ggcgctgcca ctgcgttcgt agatcgtctt gaacaaccat ctggcttctg
     8701 ccttgcctgc ggcgcggcgt gccaggcggt agagaaaacg gccgatgccg ggatcgatca
     8761 aaaagtaatc ggggtgaacc gtcagcacgt ccgggttctt gccttctgtg atctcgcggt
     8821 acatccaatc agctagctcg atctcgatgt actccggccg cccggtttcg ctctttacga
     8881 tcttgtagcg gctaatcaag gcttcaccct cggataccgt caccaggcgg ccgttcttgg
     8941 ccttcttcgt acgctgcatg gcaacgtgcg tggtgtttaa ccgaatgcag gtttctacca
     9001 ggtcgtcttt ctgctttccg ccatcggctc gccggcagaa cttgagtacg tccgcaacgt
     9061 gtggacggaa cacgcggccg ggcttgtctc ccttcccttc ccggtatcgg ttcatggatt
     9121 cggttagatg ggaaaccgcc atcagtacca ggtcgtaatc ccacacactg gccatgccgg
     9181 ccggccctgc ggaaacctct acgtgcccgt ctggaagctc gtagcggatc acctcgccag
     9241 ctcgtcggtc acgcttcgac agacggaaaa cggccacgtc catgatgctg cgactatcgc
     9301 gggtgcccac gtcatagagc atcggaacga aaaaatctgg ttgctcgtcg cccttgggcg
     9361 gcttcctaat cgacgacgca ccggctgccg gcggttgccg ggattctttg cggattcgat
     9421 cagcggccgc ttgccacgat tcaccggggc gtgcttctgc ctcgatgcgt tgccgctggg
     9481 cggcctgcgc ggccttcaac ttctccacca ggtcatcacc cagcgccgcg ccgatttgta
     9541 ccgggccgga tggtttgcga ccgctcacgc cgattcctcg ggcttggggg ttccagtgcc
     9601 attgcagggc cggcagacaa cccagccgct tacgcctggc caaccgcccg ttcctccaca
     9661 catggggcat tccacggcgt cggtgcctgg ttgttcttga ttttccatgc cgcctccttt
     9721 agcc
//
