LOCUS       sequence-342805-        8353 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 8353)
  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 8353)
  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_14371
FEATURES             Location/Qualifiers
     source          1..8353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    757..781
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(848..1555)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1923..2268)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1960..1980)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3301..3553)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4153..4335)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4189..4367)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4530..4554
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4690..5278)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4770..4789)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5449..6243)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6244..6348)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6410..6604
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6670..7743)
                     /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(8172..8353,1..448))
                     /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 cgccgatgcg cgtgcctttg atcgcccgcg acacgacaaa ggccgcttgt agccttccat
       61 ccgtgacctc aatgcgctgc ttaaccagct ccaccaggtc ggcggtggcc catatgtcgt
      121 aagggcttgg ctgcaccgga atcagcacga agtcggctgc cttgatcgcg gacacagcca
      181 agtccgccgc ctggggcgct ccgtcgatca ctacgaagtc gcgccggccg atggccttca
      241 cgtcgcggtc aatcgtcggg cggtcgatgc cgacaacggt tagcggttga tcttcccgca
      301 cggccgccca atcgcgggca ctgccctggg gatcggaatc gactaacaga acatcggccc
      361 cggcgagttg cagggcgcgg gctagatggg ttgcgatggt cgtcttgcct gacccgcctt
      421 tctggttaag tacagcgata accttcatgc gttccccttg cgtatttgtt tatttactca
      481 tcgcatcata tacgcagcga ccgcatgacg caagctgttt tactcaaata cacatcacct
      541 ttttagacgg cggcgctcgg tttcttcagc ggccaagctg gccggccagg ccgccagctt
      601 ggcatcagac aaaccggcca ggatttcatg cagccgcacg gctgactccc gaggggaacc
      661 ctgtggttgg catgcacata caaatggacg aacggataaa ccttttcacg cccttttaaa
      721 tatccgatta ttctaataaa cgctcttttc tcttaggttt acccgccaat atatcctgtc
      781 aaacactgat agtttaaacg gatcacttcg tgtccctcgg tcacatgtgc atcctctagt
      841 agcggtcctg ctgagcctcg acatgttgtc gcaaaattcg ccctggaccc gcccaacgat
      901 ttgtcgtcac tgtcaaggtt tgacctgcac ttcatttggg gcccacatac accaaaaaaa
      961 tgctgcataa ttctcggggc agcaagtcgg ttacccggcc gccgtgctgg accgggttga
     1021 atggtgcccg taactttcgg tagagcggac ggccaatact caacttcaag gaatctcacc
     1081 catgcgcgcc ggcggggaac cggagttccc ttcagtgagc gttattagtt cgccgctcgg
     1141 tgtgtcgtag atactagccc ctggggcact tttgaaattt gaataagatt tatgtaatca
     1201 gtcttttagg tttgaccggt tctgccgctt tttttaaaat tggatttgta ataataaaac
     1261 gcaattgttt gttattgtgg cgctctatca tagatgtcgc tataaaccta ttcagcacaa
     1321 tatattgttt tcattttaat attgtacata taagtagtag ggtacaatca gtaaattgaa
     1381 cggagaatat tattcataaa aatacgatag taacgggtga tatattcatt agaatgaacc
     1441 gaaaccggcg gtaaggatct gagctacaca tgctcaggtt ttttacaacg tgcacaacag
     1501 aattgaaagc aaatatcatg cgatcatagg cttctcgcat atctcattaa agcaggacaa
     1561 gctcagtatc cagtcaatcc caagtgcatc atgtaagagt tgtagatctg gttgtactta
     1621 gctccctgag tctttctaga agccaacttc aagaatgtgt cgatctggtc caaagtcacc
     1681 ctgttgtgct gccaccacat gtaagcagcg tcagccaact tgtcgtttcc catcaactgc
     1741 tttgcaagct tctttgcgtc agccctctca gtcaaagcag ccacgttgaa catctcctcg
     1801 ttcaagttag ccaagttgaa gttaggagcc attgtatcga taattgtaaa tgtaattgta
     1861 atgttgtttg ttgtttgttg ttgttggtaa ttgttgtaaa aatgagctct tatactcgag
     1921 cgtgtcctct ccaaatgaaa tgaacttcct tatatagagg aagggtcttg cgaaggatag
     1981 tgggattgtg cgtcatccct tacgtcagtg gagatgtcac atcaatccac ttgctttgta
     2041 gacgtggttg gaacctcttc tttttccacg atgctcctcg tgggtggggg tccatctttg
     2101 ggaccactgt cggcagagag atcttgaatg atagcctttc ctttatcgca atgatggcat
     2161 ttgtaggagc caccttcctt ttctactgtc ctttcgatga agtgacagat agctgggcaa
     2221 tggaatccga ggaggtttcc cgaaattatc ctttgttgaa aagtctcaat agccctttga
     2281 tcttctgaga ctgtatcttt gacatttttg gagtagacca gagtgtcgtg ctccaccatg
     2341 ttgacgaaga ttttcttctt gtcattgagt cgtaaaagac tctgtatgaa ctgttcgcca
     2401 gtcttgacgg cgagttctgt tagatcctcg atttgaatct tagactccat gcatggcctt
     2461 agattcagta ggaactacct ttttagagac tccaatctct attacttgcc ttggtttatg
     2521 aagcaagcct tgaatcgtcc atactggaat agtacttctg atcttgagaa atatgtcttt
     2581 ctctgtgttc ttgatgcaat tagtcctgaa tcttttgact gcatctttaa ccttcttggg
     2641 aaggtatttg atctcctgga gattgttact cgggtagatc gtcttgatga gacttgctgc
     2701 gtaggcctct ctaaccatct gtgggtcagc attctttctg aaattgaaga ggctaacctt
     2761 ctcattatca gtggtgaaca tagtgtcgtc accttcacct tcgaacttcc ttcctagatc
     2821 gtaaagatag aggaaatcgt ccattgtaat ctccggggca aaggagatct cttttggggc
     2881 tggatcactg ctgggccttt tggttcctag cgtgagccag tgggcttttt gctttggtgg
     2941 gcttgttagg gccttagcaa agctcttggg cttgagttga gcttctcctt tggggatgaa
     3001 gttcaacctg tctgtttgct gacttgttgt gtacgcgtca gctgctgctc ttgcctctgt
     3061 aatagtggca aatttcttgt gtgcaactcc gggaacgccg tttgttgccg cctttgtaca
     3121 accccagtca tcgtatatac cggcatgtgg accgttatac acaacgtagt agttgatatg
     3181 agggtgttga atacccgatt ctgctctgag aggagcaact gtgctgttaa gctcagattt
     3241 ttgtgggatt ggaattcctc caagcttgaa ttcttgcagc gctccaattc agcaagcgtc
     3301 gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt
     3361 tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca
     3421 taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt
     3481 atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa
     3541 tgtttgaacg atctgcttga ctctagatcc agagtcccgc tcaaatctca gtcacaggaa
     3601 gaacaggtct aggaggaaca ggaagagaga aatcaagctg ccagaatccc acatcatgcc
     3661 aatttccatg tttgaaacca gcagctctaa gcattcccct aggagcataa ccaagagcct
     3721 catgcatcct cacagaagga tcattaggaa gtccaatcac agcaaccaca gacttgaaac
     3781 cctgagcctc caaagacttc aaaagatgag tgtaaagagt agatccaaga ccagttcttt
     3841 gatgcctagg agacacgtaa acagtagact cagcagtcca atcgtaagca ttcctagcct
     3901 tccaaggacc agcataagca ataccagcca cttcaccatc cacttcagca acaagccaag
     3961 gatacctctc tctaagcctc acaagatcat cagtccactc ttgtggttct tgaggctcag
     4021 tcctgaaatt cacagtagaa gtctcaatgt aatgattcac aatagtgcac acagcaggca
     4081 tatcagcctc agtagccctt ctaatatcag caggccttct ttcaggagac atgcgaaacg
     4141 atccagatcc ggtgcagatt atttggattg agagtgaata tgagactcta attggatacc
     4201 gaggggaatt tatggaacgt cagtggagca ttttgacaag aaatatttgc tagctgatag
     4261 tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc ttagctcatt
     4321 aaactccaga aacccgcggc tgagtggctc cttcaacgtt gcggttctgt cagttccaaa
     4381 cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc ttaattctcc
     4441 gctccggatc cgaattcggc acagagtgtt caaccccagt acattaaaaa cgtccgcaat
     4501 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
     4561 agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca
     4621 tcaaccatga ctactgtccg gtccaacaga tccgagcagg aaccgtaaaa aggccgcgtt
     4681 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4741 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4801 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4861 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4921 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4981 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     5041 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     5101 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     5161 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     5221 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5281 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5341 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5401 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct aaaacaattc
     5461 atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa
     5521 aaatagctcg acatactgtt cttccccgat atcctccctg agcgaccgga cgcagaaggc
     5581 aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt
     5641 gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc
     5701 ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc
     5761 ctcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc
     5821 ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa
     5881 gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata
     5941 ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg
     6001 ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc
     6061 cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata
     6121 taggttttca ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc
     6181 ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc
     6241 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     6301 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca ctcggagcct
     6361 tgatgtgggc gctttttgca gctgttcggc tgtgcgctgg ccagacagtt atgcacaggc
     6421 caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt
     6481 tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt
     6541 cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc
     6601 cacaggaaag aagctttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg
     6661 ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac
     6721 taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga
     6781 cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg
     6841 ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag
     6901 gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag
     6961 cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc
     7021 gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc
     7081 accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac
     7141 gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc
     7201 ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt
     7261 gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag
     7321 taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg
     7381 cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg
     7441 gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg
     7501 aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg acgcaccggc
     7561 tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc
     7621 ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc
     7681 caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct
     7741 cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca gacaacccag
     7801 ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg
     7861 cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca tctactcatt
     7921 tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca gctcggtaat
     7981 ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg ccggcaactg
     8041 aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg cagccttgct
     8101 gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca ttttctcttt
     8161 acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg gacctcgcgg
     8221 gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc agtgcctggg
     8281 tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc ggccagcgcc
     8341 tcggcaacct cac
//
