LOCUS       sequence-342350-        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_15125
FEATURES             Location/Qualifiers
     source          1..9484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    766..790
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(857..1564)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3063..3408)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3100..3120)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4441..4693)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5293..5475)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5329..5507)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5670..5694
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5830..6418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5910..5929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6589..7383)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7384..7488)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7550..7744
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7810..8883)
                     /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"
     CDS             complement(join(9312..9484,1..457))
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 caacctcacc gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta
       61 gccttccatc cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc
      121 atatgtcgta agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg
      181 acacagccaa gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg cgccggccga
      241 tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat
      301 cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg actaacagaa
      361 catcggcccc ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg
      421 acccgccttt ctggttaagt acagcgataa ccttcatgcg ttccccttgc gtatttgttt
      481 atttactcat cgcatcatat acgcagcgac cgcatgacgc aagctgtttt actcaaatac
      541 acatcacctt tttagacggc ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc
      601 cgccagcttg gcatcagaca aaccggccag gatttcatgc agccgcacgg ctgactcccg
      661 aggggaaccc tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc
      721 ccttttaaat atccgattat tctaataaac gctcttttct cttaggttta cccgccaata
      781 tatcctgtca aacactgata gtttaaacgg atcacttcgt gtccctcggt cacatgtgca
      841 tcctctagta gcggtcctgc tgagcctcga catgttgtcg caaaattcgc cctggacccg
      901 cccaacgatt tgtcgtcact gtcaaggttt gacctgcact tcatttgggg cccacataca
      961 ccaaaaaaat gctgcataat tctcggggca gcaagtcggt tacccggccg ccgtgctgga
     1021 ccgggttgaa tggtgcccgt aactttcggt agagcggacg gccaatactc aacttcaagg
     1081 aatctcaccc atgcgcgccg gcggggaacc ggagttccct tcagtgagcg ttattagttc
     1141 gccgctcggt gtgtcgtaga tactagcccc tggggcactt ttgaaatttg aataagattt
     1201 atgtaatcag tcttttaggt ttgaccggtt ctgccgcttt ttttaaaatt ggatttgtaa
     1261 taataaaacg caattgtttg ttattgtggc gctctatcat agatgtcgct ataaacctat
     1321 tcagcacaat atattgtttt cattttaata ttgtacatat aagtagtagg gtacaatcag
     1381 taaattgaac ggagaatatt attcataaaa atacgatagt aacgggtgat atattcatta
     1441 gaatgaaccg aaaccggcgg taaggatctg agctacacat gctcaggttt tttacaacgt
     1501 gcacaacaga attgaaagca aatatcatgc gatcataggc ttctcgcata tctcattaaa
     1561 gcaggacaag ctcacctcct agggttgata gaagtgtcga tccttggtgg tgccatgtgc
     1621 tctttgtagt agtccttgta cctagctgca gtagccctct ccaagtttga agcagtgttc
     1681 tcagaagatc tgaagatagt accgtagatg tcagatggct ctgtctccct cttgaaccac
     1741 cttccgaaca acatcctctg gtattcagtt gcgatagaga tctcgtccaa gtcagcgtaa
     1801 ccgttcctga taaggtgctc cctagccctt gcagtctgga tagccaactc gtcctcacca
     1861 ccgaaaccct ttgacacagc cctcaacatt ccctcgttgt acaacaagtt cttgttagac
     1921 aaagtagcca agtatcctga caaagtgtcg aacttaggat tagtaggaga caaaaggtcg
     1981 ttgaaatgga tgtcaagaag cctgaacaca gagtttgctg agattccacc ctccttccac
     2041 ttcaagaact gctgcctctg caatgtgatc accaagttca tagttgcagc gtcgtctggc
     2101 ttagcgaaag tcaaagcagt tgccactgct ttatttccgt actgagccac aaatgagtcg
     2161 atcaaagatg cctttgaatt tgggtttttt gcgttataca ttgagatgaa agctccgaaa
     2221 gtgtgcacgt tcctgtcaag caatgcgtcc aatcccctgt tcaacctcaa ctttttgtag
     2281 atatgtttag gtctaagaag ctccaccctc acccacttgt tgataagagc gttctgcaaa
     2341 gctgaagcaa gctcccttgt cttaacgtcc tcccttgctc tagcaaggat tgttgcaagc
     2401 tgctcctctc caccaaatcc tttagtcaaa gttttcaaaa gtgagtcttt agtagacttt
     2461 gttgaagtaa ccaatttaat gtagtcgtca agcacttcca tcttaggaga agccaatgcc
     2521 aaatatccat cagacctcaa tttaagcaaa gtaaaaacgt catcaactga tttcctcttc
     2581 tgcaaccaag aaagcaactg agcgtgccta agcctctgga tagtgttcat ctcaccacca
     2641 gccctctgtg cagtcatcaa tgcgtttgcc acagcgtcgt caccgtagtg agccatgaac
     2701 ataccgatca ctgaagcctt gttgttgttg ttctgagtat tcaactcttt aacataattt
     2761 tccaatgcct taaggttagg tgaagcaaga gcgtcgtcca ttccgtttgt caacttgaac
     2821 ctcctcaaga taggctcagc tggagggtcc ttagagatcc atgatccaag gttgatctta
     2881 gacactccag ccttagcaag gtctgtaagc ttctccaatc ctgctgagat caccctgttc
     2941 tcaccattct cagtgttctg tgtagacacc attgtatcga taattgtaaa tgtaattgta
     3001 atgttgtttg ttgtttgttg ttgttggtaa ttgttgtaaa aatgagctct tatactcgag
     3061 cgtgtcctct ccaaatgaaa tgaacttcct tatatagagg aagggtcttg cgaaggatag
     3121 tgggattgtg cgtcatccct tacgtcagtg gagatgtcac atcaatccac ttgctttgta
     3181 gacgtggttg gaacctcttc tttttccacg atgctcctcg tgggtggggg tccatctttg
     3241 ggaccactgt cggcagagag atcttgaatg atagcctttc ctttatcgca atgatggcat
     3301 ttgtaggagc caccttcctt ttctactgtc ctttcgatga agtgacagat agctgggcaa
     3361 tggaatccga ggaggtttcc cgaaattatc ctttgttgaa aagtctcaat agccctttga
     3421 tcttctgaga ctgtatcttt gacatttttg gagtagacca gagtgtcgtg ctccaccatg
     3481 ttgacgaaga ttttcttctt gtcattgagt cgtaaaagac tctgtatgaa ctgttcgcca
     3541 gtcttgacgg cgagttctgt tagatcctcg atttgaatct tagactccat gcatggcctt
     3601 agattcagta ggaactacct ttttagagac tccaatctct attacttgcc ttggtttatg
     3661 aagcaagcct tgaatcgtcc atactggaat agtacttctg atcttgagaa atatgtcttt
     3721 ctctgtgttc ttgatgcaat tagtcctgaa tcttttgact gcatctttaa ccttcttggg
     3781 aaggtatttg atctcctgga gattgttact cgggtagatc gtcttgatga gacttgctgc
     3841 gtaggcctct ctaaccatct gtgggtcagc attctttctg aaattgaaga ggctaacctt
     3901 ctcattatca gtggtgaaca tagtgtcgtc accttcacct tcgaacttcc ttcctagatc
     3961 gtaaagatag aggaaatcgt ccattgtaat ctccggggca aaggagatct cttttggggc
     4021 tggatcactg ctgggccttt tggttcctag cgtgagccag tgggcttttt gctttggtgg
     4081 gcttgttagg gccttagcaa agctcttggg cttgagttga gcttctcctt tggggatgaa
     4141 gttcaacctg tctgtttgct gacttgttgt gtacgcgtca gctgctgctc ttgcctctgt
     4201 aatagtggca aatttcttgt gtgcaactcc gggaacgccg tttgttgccg cctttgtaca
     4261 accccagtca tcgtatatac cggcatgtgg accgttatac acaacgtagt agttgatatg
     4321 agggtgttga atacccgatt ctgctctgag aggagcaact gtgctgttaa gctcagattt
     4381 ttgtgggatt ggaattcctc caagcttgaa ttcttgcagc gctccaattc agcaagcgtc
     4441 gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt
     4501 tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca
     4561 taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt
     4621 atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa
     4681 tgtttgaacg atctgcttga ctctagatcc agagtcccgc tcaaatctca gtcacaggaa
     4741 gaacaggtct aggaggaaca ggaagagaga aatcaagctg ccagaatccc acatcatgcc
     4801 aatttccatg tttgaaacca gcagctctaa gcattcccct aggagcataa ccaagagcct
     4861 catgcatcct cacagaagga tcattaggaa gtccaatcac agcaaccaca gacttgaaac
     4921 cctgagcctc caaagacttc aaaagatgag tgtaaagagt agatccaaga ccagttcttt
     4981 gatgcctagg agacacgtaa acagtagact cagcagtcca atcgtaagca ttcctagcct
     5041 tccaaggacc agcataagca ataccagcca cttcaccatc cacttcagca acaagccaag
     5101 gatacctctc tctaagcctc acaagatcat cagtccactc ttgtggttct tgaggctcag
     5161 tcctgaaatt cacagtagaa gtctcaatgt aatgattcac aatagtgcac acagcaggca
     5221 tatcagcctc agtagccctt ctaatatcag caggccttct ttcaggagac atgcgaaacg
     5281 atccagatcc ggtgcagatt atttggattg agagtgaata tgagactcta attggatacc
     5341 gaggggaatt tatggaacgt cagtggagca ttttgacaag aaatatttgc tagctgatag
     5401 tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc ttagctcatt
     5461 aaactccaga aacccgcggc tgagtggctc cttcaacgtt gcggttctgt cagttccaaa
     5521 cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc ttaattctcc
     5581 gctccggatc cgaattcggc acagagtgtt caaccccagt acattaaaaa cgtccgcaat
     5641 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
     5701 agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca
     5761 tcaaccatga ctactgtccg gtccaacaga tccgagcagg aaccgtaaaa aggccgcgtt
     5821 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     5881 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     5941 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6001 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6061 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6121 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6181 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6241 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6301 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6361 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     6421 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6481 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6541 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct aaaacaattc
     6601 atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa
     6661 aaatagctcg acatactgtt cttccccgat atcctccctg agcgaccgga cgcagaaggc
     6721 aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt
     6781 gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc
     6841 ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc
     6901 ctcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc
     6961 ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa
     7021 gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata
     7081 ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg
     7141 ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc
     7201 cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata
     7261 taggttttca ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc
     7321 ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc
     7381 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     7441 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca ctcggagcct
     7501 tgatgtgggc gctttttgca gctgttcggc tgtgcgctgg ccagacagtt atgcacaggc
     7561 caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt
     7621 tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt
     7681 cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc
     7741 cacaggaaag aagctttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg
     7801 ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac
     7861 taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga
     7921 cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg
     7981 ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag
     8041 gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag
     8101 cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc
     8161 gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc
     8221 accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac
     8281 gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc
     8341 ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt
     8401 gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag
     8461 taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg
     8521 cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg
     8581 gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg
     8641 aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg acgcaccggc
     8701 tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc
     8761 ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc
     8821 caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct
     8881 cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca gacaacccag
     8941 ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg
     9001 cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca tctactcatt
     9061 tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca gctcggtaat
     9121 ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg ccggcaactg
     9181 aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg cagccttgct
     9241 gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca ttttctcttt
     9301 acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg gacctcgcgg
     9361 gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc agtgcctggg
     9421 tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc ggccagcgcc
     9481 tcgg
//
