LOCUS       sequence-342197-        9304 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 9304)
  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 9304)
  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_06478a
FEATURES             Location/Qualifiers
     source          1..9304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(298..927)
                     /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    1236..1260
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1327..2034)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(2325..2336)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(3353..3698)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3390..3410)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4731..4983)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5583..5765)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5619..5797)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5960..5984
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6120..6708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6200..6219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6879..7673)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7674..7778)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7840..8034
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8100..9173)
                     /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 attttccatg ccgcctcctt tagccgctaa aattcatcta ctcatttatt catttgctca
       61 tttactctgg tagctgcgcg atgtattcag atagcagctc ggtaatggtc ttgccttggc
      121 gtaccgcgta catcttcagc ttggtgtgat cctccgccgg caactgaaag ttgacccgct
      181 tcatggctgg cgtgtctgcc aggctggcca acgttgcagc cttgctgctg cgtgcgctcg
      241 gacggccggc acttagcgtg tttgtgcttt tgctcatttt ctctttacct cattaactca
      301 aatgagtttt gatttaattt cagcggccag cgcctggacc tcgcgggcag cgtcgccctc
      361 gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg
      421 cgtgatacgg gactcaagaa tgggcagctc gtacccggcc agcgcctcgg caacctcacc
      481 gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta gccttccatc
      541 cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc atatgtcgta
      601 agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa
      661 gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg cgccggccga tggccttcac
      721 gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat cttcccgcac
      781 ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg actaacagaa catcggcccc
      841 ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg acccgccttt
      901 ctggttaagt acagcgataa ccttcatgcg ttccccttgc gtatttgttt atttactcat
      961 cgcatcatat acgcagcgac cgcatgacgc aagctgtttt actcaaatac acatcacctt
     1021 tttagacggc ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc cgccagcttg
     1081 gcatcagaca aaccggccag gatttcatgc agccgcacgg ctgactcccg aggggaaccc
     1141 tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc ccttttaaat
     1201 atccgattat tctaataaac gctcttttct cttaggttta cccgccaata tatcctgtca
     1261 aacactgata gtttaaacgg atcacttcgt gtccctcggt cacatgtgca tcctctagta
     1321 gcggtcctgc tgagcctcga catgttgtcg caaaattcgc cctggacccg cccaacgatt
     1381 tgtcgtcact gtcaaggttt gacctgcact tcatttgggg cccacataca ccaaaaaaat
     1441 gctgcataat tctcggggca gcaagtcggt tacccggccg ccgtgctgga ccgggttgaa
     1501 tggtgcccgt aactttcggt agagcggacg gccaatactc aacttcaagg aatctcaccc
     1561 atgcgcgccg gcggggaacc ggagttccct tcagtgagcg ttattagttc gccgctcggt
     1621 gtgtcgtaga tactagcccc tggggcactt ttgaaatttg aataagattt atgtaatcag
     1681 tcttttaggt ttgaccggtt ctgccgcttt ttttaaaatt ggatttgtaa taataaaacg
     1741 caattgtttg ttattgtggc gctctatcat agatgtcgct ataaacctat tcagcacaat
     1801 atattgtttt cattttaata ttgtacatat aagtagtagg gtacaatcag taaattgaac
     1861 ggagaatatt attcataaaa atacgatagt aacgggtgat atattcatta gaatgaaccg
     1921 aaaccggcgg taaggatctg agctacacat gctcaggttt tttacaacgt gcacaacaga
     1981 attgaaagca aatatcatgc gatcataggc ttctcgcata tctcattaaa gcaggacaag
     2041 ctcattgtgc cttcatagct gcagtaagag ctccgattgg gttatcgttg tcaagaagtg
     2101 ttttaacctt gttaagcaaa gacactcctc ccttcctctc tgtgtagtac aatgtgtaag
     2161 caacccaagt tttgtaaatt gggtcgttct taggcaaata cacagcagaa gtagggaatg
     2221 ggatagcaag catagatgac acagccttag ggtcccactc agtgtgcaac caattctgga
     2281 acaaataagt ttgcatgttc tcagcaagcc tcttaaggtc aggcacgtcc ttcaatgact
     2341 caaagaatgt agcgaactgc cactctgacc tcaatggctt tgtgtcagca aggaagtgca
     2401 ccatctgcac agatgacaac acgttcttct ttgtgtgaga tctgtacaaa ttaacgtacc
     2461 taagccaatg gatgaacatt gcgtttgtgt ccacagcgtc catgttcacc ttcacaagcc
     2521 taaggatttt gaacacctgc tcaggtgtca cgtcgtaatt caaccacacg tcgtgcatca
     2581 tcttcctaga agtctttgaa gaaaggaaca agaacctctg cattgtgtca ccgtggtcct
     2641 tcattccagg cacgttcctc aaaaggtgca acaatgtcac cacgtccttc tctggcattg
     2701 tttttctgaa aaggtcaagc atgtcgtcgt caccgaacca gtggtttccc ttctgagccc
     2761 tgtactgctg agcgtatgcc aaccactggt ggaagattgg agagtccacc aactttgttc
     2821 cagcctcctg caacctcaat gtatcgtaca caatcttagg accagtctcg tgctgcaaag
     2881 ttttagacac ccatgaacca gacaaagatt tttgcataga ctttcccaac tccttaagag
     2941 ctcctgtctt tctgaatgac tcgatcaaga tcttcaattc ctctcctgaa gatctcttca
     3001 acaacaagta gtacaagtca aggtctgaga aagcctggtt tccgttcttt gccctgaaag
     3061 cagccaccaa cttaagccac aacaacaaat taggatcgtc gttaagcttt gtagtagatt
     3121 ttgccaatct caatgtgtcg aaaatagcct gagggtgcac cctagacttc atccaagcct
     3181 tgatctgagc ctgctcaggg ttagacttga acaccttatg gataagatcg tcaaccttag
     3241 acaaaagtgg agtagttccc attgtatcga taattgtaaa tgtaattgta atgttgtttg
     3301 ttgtttgttg ttgttggtaa ttgttgtaaa aatgagctct tatactcgag cgtgtcctct
     3361 ccaaatgaaa tgaacttcct tatatagagg aagggtcttg cgaaggatag tgggattgtg
     3421 cgtcatccct tacgtcagtg gagatgtcac atcaatccac ttgctttgta gacgtggttg
     3481 gaacctcttc tttttccacg atgctcctcg tgggtggggg tccatctttg ggaccactgt
     3541 cggcagagag atcttgaatg atagcctttc ctttatcgca atgatggcat ttgtaggagc
     3601 caccttcctt ttctactgtc ctttcgatga agtgacagat agctgggcaa tggaatccga
     3661 ggaggtttcc cgaaattatc ctttgttgaa aagtctcaat agccctttga tcttctgaga
     3721 ctgtatcttt gacatttttg gagtagacca gagtgtcgtg ctccaccatg ttgacgaaga
     3781 ttttcttctt gtcattgagt cgtaaaagac tctgtatgaa ctgttcgcca gtcttgacgg
     3841 cgagttctgt tagatcctcg atttgaatct tagactccat gcatggcctt agattcagta
     3901 ggaactacct ttttagagac tccaatctct attacttgcc ttggtttatg aagcaagcct
     3961 tgaatcgtcc atactggaat agtacttctg atcttgagaa atatgtcttt ctctgtgttc
     4021 ttgatgcaat tagtcctgaa tcttttgact gcatctttaa ccttcttggg aaggtatttg
     4081 atctcctgga gattgttact cgggtagatc gtcttgatga gacttgctgc gtaggcctct
     4141 ctaaccatct gtgggtcagc attctttctg aaattgaaga ggctaacctt ctcattatca
     4201 gtggtgaaca tagtgtcgtc accttcacct tcgaacttcc ttcctagatc gtaaagatag
     4261 aggaaatcgt ccattgtaat ctccggggca aaggagatct cttttggggc tggatcactg
     4321 ctgggccttt tggttcctag cgtgagccag tgggcttttt gctttggtgg gcttgttagg
     4381 gccttagcaa agctcttggg cttgagttga gcttctcctt tggggatgaa gttcaacctg
     4441 tctgtttgct gacttgttgt gtacgcgtca gctgctgctc ttgcctctgt aatagtggca
     4501 aatttcttgt gtgcaactcc gggaacgccg tttgttgccg cctttgtaca accccagtca
     4561 tcgtatatac cggcatgtgg accgttatac acaacgtagt agttgatatg agggtgttga
     4621 atacccgatt ctgctctgag aggagcaact gtgctgttaa gctcagattt ttgtgggatt
     4681 ggaattcctc caagcttgaa ttcttgcagc gctccaattc agcaagcgtc gatctagtaa
     4741 catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat
     4801 cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca taaataacgt
     4861 catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt atatgataat
     4921 catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa tgtttgaacg
     4981 atctgcttga ctctagatcc agagtcccgc tcaaatctca gtcacaggaa gaacaggtct
     5041 aggaggaaca ggaagagaga aatcaagctg ccagaatccc acatcatgcc aatttccatg
     5101 tttgaaacca gcagctctaa gcattcccct aggagcataa ccaagagcct catgcatcct
     5161 cacagaagga tcattaggaa gtccaatcac agcaaccaca gacttgaaac cctgagcctc
     5221 caaagacttc aaaagatgag tgtaaagagt agatccaaga ccagttcttt gatgcctagg
     5281 agacacgtaa acagtagact cagcagtcca atcgtaagca ttcctagcct tccaaggacc
     5341 agcataagca ataccagcca cttcaccatc cacttcagca acaagccaag gatacctctc
     5401 tctaagcctc acaagatcat cagtccactc ttgtggttct tgaggctcag tcctgaaatt
     5461 cacagtagaa gtctcaatgt aatgattcac aatagtgcac acagcaggca tatcagcctc
     5521 agtagccctt ctaatatcag caggccttct ttcaggagac atgcgaaacg atccagatcc
     5581 ggtgcagatt atttggattg agagtgaata tgagactcta attggatacc gaggggaatt
     5641 tatggaacgt cagtggagca ttttgacaag aaatatttgc tagctgatag tgaccttagg
     5701 cgacttttga acgcgcaata atggtttctg acgtatgtgc ttagctcatt aaactccaga
     5761 aacccgcggc tgagtggctc cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg
     5821 cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc ttaattctcc gctccggatc
     5881 cgaattcggc acagagtgtt caaccccagt acattaaaaa cgtccgcaat gtgttattaa
     5941 gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc agccaacagc
     6001 tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca tcaaccatga
     6061 ctactgtccg gtccaacaga tccgagcagg aaccgtaaaa aggccgcgtt gctggcgttt
     6121 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6181 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6241 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6301 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6361 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6421 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6481 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     6541 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     6601 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     6661 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     6721 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     6781 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     6841 tcaatctaaa gtatatatga gtaaacttgg tctgacagct aaaacaattc atccagtaaa
     6901 atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg
     6961 acatactgtt cttccccgat atcctccctg agcgaccgga cgcagaaggc aatgtcatac
     7021 cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc
     7081 acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt
     7141 tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ctcttcccag
     7201 ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg
     7261 ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg
     7321 cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata ctcttccgag
     7381 caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag
     7441 tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt
     7501 tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata taggttttca
     7561 ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag
     7621 cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc catactcttc
     7681 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7741 gaatgtattt agaaaaataa acaaataggg gttccgcgca ctcggagcct tgatgtgggc
     7801 gctttttgca gctgttcggc tgtgcgctgg ccagacagtt atgcacaggc caggcgggtt
     7861 ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt tttctctttt
     7921 atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt cccaatgtac
     7981 gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc cacaggaaag
     8041 aagctttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg ctccgtacat
     8101 taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac taggatcggg
     8161 ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga cccgatcagc
     8221 ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg ttcgtagatc
     8281 gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag gcggtagaga
     8341 aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag cacgtccggg
     8401 ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc gatgtactcc
     8461 ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc accctcggat
     8521 accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac gtgcgtggtg
     8581 tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc ggctcgccgg
     8641 cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt gtctcccttc
     8701 ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag taccaggtcg
     8761 taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg cccgtctgga
     8821 agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg gaaaacggcc
     8881 acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg aacgaaaaaa
     8941 tctggttgct cgtcgccctt gggcggcttc ctaatcgacg acgcaccggc tgccggcggt
     9001 tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc ggggcgtgct
     9061 tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc caccaggtca
     9121 tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct cacgccgatt
     9181 cctcgggctt gggggttcca gtgccattgc agggccggca gacaacccag ccgcttacgc
     9241 ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg cctggttgtt
     9301 cttg
//
