LOCUS       sequence-342452-        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_15127a
FEATURES             Location/Qualifiers
     source          1..9484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(68..697)
                     /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    1006..1030
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1097..1804)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3303..3648)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3340..3360)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4681..4933)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5533..5715)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5569..5747)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5910..5934
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6070..6658)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6150..6169)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6829..7623)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7624..7728)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7790..7984
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8050..9123)
                     /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 cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt tgctcatttt ctctttacct
       61 cattaactca aatgagtttt gatttaattt cagcggccag cgcctggacc tcgcgggcag
      121 cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc
      181 tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc gtacccggcc agcgcctcgg
      241 caacctcacc gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta
      301 gccttccatc cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc
      361 atatgtcgta agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg
      421 acacagccaa gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg cgccggccga
      481 tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat
      541 cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg actaacagaa
      601 catcggcccc ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg
      661 acccgccttt ctggttaagt acagcgataa ccttcatgcg ttccccttgc gtatttgttt
      721 atttactcat cgcatcatat acgcagcgac cgcatgacgc aagctgtttt actcaaatac
      781 acatcacctt tttagacggc ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc
      841 cgccagcttg gcatcagaca aaccggccag gatttcatgc agccgcacgg ctgactcccg
      901 aggggaaccc tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc
      961 ccttttaaat atccgattat tctaataaac gctcttttct cttaggttta cccgccaata
     1021 tatcctgtca aacactgata gtttaaacgg atcacttcgt gtccctcggt cacatgtgca
     1081 tcctctagta gcggtcctgc tgagcctcga catgttgtcg caaaattcgc cctggacccg
     1141 cccaacgatt tgtcgtcact gtcaaggttt gacctgcact tcatttgggg cccacataca
     1201 ccaaaaaaat gctgcataat tctcggggca gcaagtcggt tacccggccg ccgtgctgga
     1261 ccgggttgaa tggtgcccgt aactttcggt agagcggacg gccaatactc aacttcaagg
     1321 aatctcaccc atgcgcgccg gcggggaacc ggagttccct tcagtgagcg ttattagttc
     1381 gccgctcggt gtgtcgtaga tactagcccc tggggcactt ttgaaatttg aataagattt
     1441 atgtaatcag tcttttaggt ttgaccggtt ctgccgcttt ttttaaaatt ggatttgtaa
     1501 taataaaacg caattgtttg ttattgtggc gctctatcat agatgtcgct ataaacctat
     1561 tcagcacaat atattgtttt cattttaata ttgtacatat aagtagtagg gtacaatcag
     1621 taaattgaac ggagaatatt attcataaaa atacgatagt aacgggtgat atattcatta
     1681 gaatgaaccg aaaccggcgg taaggatctg agctacacat gctcaggttt tttacaacgt
     1741 gcacaacaga attgaaagca aatatcatgc gatcataggc ttctcgcata tctcattaaa
     1801 gcaggacaag ctcacctcct tggattaatt gaagtgtcaa tccttggagg agccatgtgc
     1861 tccttgtagt agtccttgta ccttgcagca gtagccctct caaggttaga agcagtgttt
     1921 tcagatgacc taaaagtagt accgtagatg tcagatggtt ttgtctttct cttgaaccac
     1981 ctaccaaaca acattctttg gtactctgta gcgatagaga tctcgtccaa gtctgcgtaa
     2041 ccgtttctga tcaagtgctc cctagcccta gcagtctgga tagccaactc gtcctctcct
     2101 ccgaatccct ttgacacagc cctcaacata ccctcgttgt acaacaagtt cttgttagaa
     2161 agagttgcca agtatcctga caatgtgtca aattttgggt ttgtaggtga caaaagatcg
     2221 ttgaagttga tatcaagaag cctaaaaaca gaattagctg agattcctcc ctccttccac
     2281 ttaaggaact gctgcctctg caatgtgata accaagttca tagttgcagc gtcgtcaggc
     2341 ttagcgaatg tcaatgcagt tgccactgcc ttgttaccgt actgtgccac gaatgagtcg
     2401 atcaaagaag ccttagagtt agggttttta gcattgtaca tagaaataaa tgcaccaaat
     2461 gtgtgaacat ttctgtccaa caaagcgtcc aatcccctgt taagcctcaa cttcttgtag
     2521 atatgctttg gcctcaacaa gtcaactcta acccacttgt taatcaatgc tgactgaaga
     2581 gccaatgcaa gctccctagt cttaacgtcc tcccttgccc tagccaagat tgttgccaac
     2641 tgctcctttc ctccgaaacc cttagtaagt gtcttaagaa gagaatcttt tgttgactta
     2701 gttgagataa caagtttaat ataatcgtca agcacctcca tcttaggtga agccaaagcc
     2761 aagtatccgt cagacctcaa cttcaaaaga gtgaaaacgt cgtccactga cttcctcttc
     2821 tgcaaccatg aaagaagctg tgcgttcctc aacctctgga tagtgttcat ctcaccacca
     2881 gccctttgtg cagtcatcaa agcgtttgcc acagcgtcat caccgtagtg tgccatgaac
     2941 ataccgatca ctgaagcctt gttgttattg ttctgtgtat tcaactcctt cacatagttt
     3001 tcaagtgcct taaggtttgg tgaagccaat gcgtcgtcca ttccatttgt caacttgaac
     3061 ctcctaagga ttggctctgc tggtgggtcc ttagagatcc atgatcccaa gttgatgtta
     3121 gacacacctg ccttagccaa gtctgtcaac ttctccaatc cagctgagat caccctgttc
     3181 tcaccgttct cagtattctg agtagacacc attgtatcga taattgtaaa tgtaattgta
     3241 atgttgtttg ttgtttgttg ttgttggtaa ttgttgtaaa aatgagctct tatactcgag
     3301 cgtgtcctct ccaaatgaaa tgaacttcct tatatagagg aagggtcttg cgaaggatag
     3361 tgggattgtg cgtcatccct tacgtcagtg gagatgtcac atcaatccac ttgctttgta
     3421 gacgtggttg gaacctcttc tttttccacg atgctcctcg tgggtggggg tccatctttg
     3481 ggaccactgt cggcagagag atcttgaatg atagcctttc ctttatcgca atgatggcat
     3541 ttgtaggagc caccttcctt ttctactgtc ctttcgatga agtgacagat agctgggcaa
     3601 tggaatccga ggaggtttcc cgaaattatc ctttgttgaa aagtctcaat agccctttga
     3661 tcttctgaga ctgtatcttt gacatttttg gagtagacca gagtgtcgtg ctccaccatg
     3721 ttgacgaaga ttttcttctt gtcattgagt cgtaaaagac tctgtatgaa ctgttcgcca
     3781 gtcttgacgg cgagttctgt tagatcctcg atttgaatct tagactccat gcatggcctt
     3841 agattcagta ggaactacct ttttagagac tccaatctct attacttgcc ttggtttatg
     3901 aagcaagcct tgaatcgtcc atactggaat agtacttctg atcttgagaa atatgtcttt
     3961 ctctgtgttc ttgatgcaat tagtcctgaa tcttttgact gcatctttaa ccttcttggg
     4021 aaggtatttg atctcctgga gattgttact cgggtagatc gtcttgatga gacttgctgc
     4081 gtaggcctct ctaaccatct gtgggtcagc attctttctg aaattgaaga ggctaacctt
     4141 ctcattatca gtggtgaaca tagtgtcgtc accttcacct tcgaacttcc ttcctagatc
     4201 gtaaagatag aggaaatcgt ccattgtaat ctccggggca aaggagatct cttttggggc
     4261 tggatcactg ctgggccttt tggttcctag cgtgagccag tgggcttttt gctttggtgg
     4321 gcttgttagg gccttagcaa agctcttggg cttgagttga gcttctcctt tggggatgaa
     4381 gttcaacctg tctgtttgct gacttgttgt gtacgcgtca gctgctgctc ttgcctctgt
     4441 aatagtggca aatttcttgt gtgcaactcc gggaacgccg tttgttgccg cctttgtaca
     4501 accccagtca tcgtatatac cggcatgtgg accgttatac acaacgtagt agttgatatg
     4561 agggtgttga atacccgatt ctgctctgag aggagcaact gtgctgttaa gctcagattt
     4621 ttgtgggatt ggaattcctc caagcttgaa ttcttgcagc gctccaattc agcaagcgtc
     4681 gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt
     4741 tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca
     4801 taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt
     4861 atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa
     4921 tgtttgaacg atctgcttga ctctagatcc agagtcccgc tcaaatctca gtcacaggaa
     4981 gaacaggtct aggaggaaca ggaagagaga aatcaagctg ccagaatccc acatcatgcc
     5041 aatttccatg tttgaaacca gcagctctaa gcattcccct aggagcataa ccaagagcct
     5101 catgcatcct cacagaagga tcattaggaa gtccaatcac agcaaccaca gacttgaaac
     5161 cctgagcctc caaagacttc aaaagatgag tgtaaagagt agatccaaga ccagttcttt
     5221 gatgcctagg agacacgtaa acagtagact cagcagtcca atcgtaagca ttcctagcct
     5281 tccaaggacc agcataagca ataccagcca cttcaccatc cacttcagca acaagccaag
     5341 gatacctctc tctaagcctc acaagatcat cagtccactc ttgtggttct tgaggctcag
     5401 tcctgaaatt cacagtagaa gtctcaatgt aatgattcac aatagtgcac acagcaggca
     5461 tatcagcctc agtagccctt ctaatatcag caggccttct ttcaggagac atgcgaaacg
     5521 atccagatcc ggtgcagatt atttggattg agagtgaata tgagactcta attggatacc
     5581 gaggggaatt tatggaacgt cagtggagca ttttgacaag aaatatttgc tagctgatag
     5641 tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc ttagctcatt
     5701 aaactccaga aacccgcggc tgagtggctc cttcaacgtt gcggttctgt cagttccaaa
     5761 cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc ttaattctcc
     5821 gctccggatc cgaattcggc acagagtgtt caaccccagt acattaaaaa cgtccgcaat
     5881 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
     5941 agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca
     6001 tcaaccatga ctactgtccg gtccaacaga tccgagcagg aaccgtaaaa aggccgcgtt
     6061 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6121 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6181 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6241 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6301 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6361 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6421 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6481 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6541 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6601 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6661 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6721 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6781 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct aaaacaattc
     6841 atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa
     6901 aaatagctcg acatactgtt cttccccgat atcctccctg agcgaccgga cgcagaaggc
     6961 aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt
     7021 gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc
     7081 ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc
     7141 ctcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc
     7201 ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa
     7261 gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata
     7321 ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg
     7381 ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc
     7441 cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata
     7501 taggttttca ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc
     7561 ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc
     7621 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7681 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca ctcggagcct
     7741 tgatgtgggc gctttttgca gctgttcggc tgtgcgctgg ccagacagtt atgcacaggc
     7801 caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt
     7861 tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt
     7921 cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc
     7981 cacaggaaag aagctttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg
     8041 ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac
     8101 taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga
     8161 cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg
     8221 ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag
     8281 gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag
     8341 cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc
     8401 gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc
     8461 accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac
     8521 gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc
     8581 ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt
     8641 gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag
     8701 taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg
     8761 cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg
     8821 gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg
     8881 aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg acgcaccggc
     8941 tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc
     9001 ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc
     9061 caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct
     9121 cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca gacaacccag
     9181 ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg
     9241 cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca tctactcatt
     9301 tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca gctcggtaat
     9361 ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg ccggcaactg
     9421 aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg cagccttgct
     9481 gctg
//
