LOCUS       sequence-342803-        8641 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 8641)
  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 8641)
  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_07558
FEATURES             Location/Qualifiers
     source          1..8641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    642..666
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(733..1440)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2096..2441)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2133..2153)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3474..3726)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4326..4508)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4362..4540)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4703..4727
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4863..5451)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4943..4962)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5622..6416)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6417..6521)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6583..6777
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6843..7916)
                     /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(8345..8641,1..333))
                     /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 gtcgtaaggg cttggctgca ccggaatcag cacgaagtcg gctgccttga tcgcggacac
       61 agccaagtcc gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc
      121 cttcacgtcg cggtcaatcg tcgggcggtc gatgccgaca acggttagcg gttgatcttc
      181 ccgcacggcc gcccaatcgc gggcactgcc ctggggatcg gaatcgacta acagaacatc
      241 ggccccggcg agttgcaggg cgcgggctag atgggttgcg atggtcgtct tgcctgaccc
      301 gcctttctgg ttaagtacag cgataacctt catgcgttcc ccttgcgtat ttgtttattt
      361 actcatcgca tcatatacgc agcgaccgca tgacgcaagc tgttttactc aaatacacat
      421 caccttttta gacggcggcg ctcggtttct tcagcggcca agctggccgg ccaggccgcc
      481 agcttggcat cagacaaacc ggccaggatt tcatgcagcc gcacggctga ctcccgaggg
      541 gaaccctgtg gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt
      601 ttaaatatcc gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc
      661 ctgtcaaaca ctgatagttt aaacggatca cttcgtgtcc ctcggtcaca tgtgcatcct
      721 ctagtagcgg tcctgctgag cctcgacatg ttgtcgcaaa attcgccctg gacccgccca
      781 acgatttgtc gtcactgtca aggtttgacc tgcacttcat ttggggccca catacaccaa
      841 aaaaatgctg cataattctc ggggcagcaa gtcggttacc cggccgccgt gctggaccgg
      901 gttgaatggt gcccgtaact ttcggtagag cggacggcca atactcaact tcaaggaatc
      961 tcacccatgc gcgccggcgg ggaaccggag ttcccttcag tgagcgttat tagttcgccg
     1021 ctcggtgtgt cgtagatact agcccctggg gcacttttga aatttgaata agatttatgt
     1081 aatcagtctt ttaggtttga ccggttctgc cgcttttttt aaaattggat ttgtaataat
     1141 aaaacgcaat tgtttgttat tgtggcgctc tatcatagat gtcgctataa acctattcag
     1201 cacaatatat tgttttcatt ttaatattgt acatataagt agtagggtac aatcagtaaa
     1261 ttgaacggag aatattattc ataaaaatac gatagtaacg ggtgatatat tcattagaat
     1321 gaaccgaaac cggcggtaag gatctgagct acacatgctc aggtttttta caacgtgcac
     1381 aacagaattg aaagcaaata tcatgcgatc ataggcttct cgcatatctc attaaagcag
     1441 gacaagctca agaagtcttt gcctccttga aaagttggta atacttgtac tcgtcgtgct
     1501 gagtcaaagc gttaccgtcc aacactctcc tagggtatcc aggcttagtc aatcccaaca
     1561 acaccttagc cacgtcgtcg tccctcctag cagatgccat gatcatagtc cttgctgtga
     1621 tctcagcctc tgaagcaccc ctagccacag ccaccactgg agggttgtat cctgcgttca
     1681 aagttgacac cacgttgtcg tcgtactgct tcacgaacct agagtaagag atgaagttct
     1741 tagtattagc aatctctttc aactgatcag ccttagtaat ctttcccaag ttcaatgact
     1801 tccaagcctc gaaagtaggc acgtggttcc tcaaccactt gttgaacaaa tagtcctcag
     1861 ccttctttgc gaagtaagcg tagtacttat agttcttgtg agccttcata gctgtgtcgt
     1921 ccaaaccgtt caacttaaga gccttcctca cgaagtcgtc tgacttctct gtctccaacc
     1981 accatctaac cttcacgtgc ttcattgtat cgataattgt aaatgtaatt gtaatgttgt
     2041 ttgttgtttg ttgttgttgg taattgttgt aaaaatgagc tcttatactc gagcgtgtcc
     2101 tctccaaatg aaatgaactt ccttatatag aggaagggtc ttgcgaagga tagtgggatt
     2161 gtgcgtcatc ccttacgtca gtggagatgt cacatcaatc cacttgcttt gtagacgtgg
     2221 ttggaacctc ttctttttcc acgatgctcc tcgtgggtgg gggtccatct ttgggaccac
     2281 tgtcggcaga gagatcttga atgatagcct ttcctttatc gcaatgatgg catttgtagg
     2341 agccaccttc cttttctact gtcctttcga tgaagtgaca gatagctggg caatggaatc
     2401 cgaggaggtt tcccgaaatt atcctttgtt gaaaagtctc aatagccctt tgatcttctg
     2461 agactgtatc tttgacattt ttggagtaga ccagagtgtc gtgctccacc atgttgacga
     2521 agattttctt cttgtcattg agtcgtaaaa gactctgtat gaactgttcg ccagtcttga
     2581 cggcgagttc tgttagatcc tcgatttgaa tcttagactc catgcatggc cttagattca
     2641 gtaggaacta cctttttaga gactccaatc tctattactt gccttggttt atgaagcaag
     2701 ccttgaatcg tccatactgg aatagtactt ctgatcttga gaaatatgtc tttctctgtg
     2761 ttcttgatgc aattagtcct gaatcttttg actgcatctt taaccttctt gggaaggtat
     2821 ttgatctcct ggagattgtt actcgggtag atcgtcttga tgagacttgc tgcgtaggcc
     2881 tctctaacca tctgtgggtc agcattcttt ctgaaattga agaggctaac cttctcatta
     2941 tcagtggtga acatagtgtc gtcaccttca ccttcgaact tccttcctag atcgtaaaga
     3001 tagaggaaat cgtccattgt aatctccggg gcaaaggaga tctcttttgg ggctggatca
     3061 ctgctgggcc ttttggttcc tagcgtgagc cagtgggctt tttgctttgg tgggcttgtt
     3121 agggccttag caaagctctt gggcttgagt tgagcttctc ctttggggat gaagttcaac
     3181 ctgtctgttt gctgacttgt tgtgtacgcg tcagctgctg ctcttgcctc tgtaatagtg
     3241 gcaaatttct tgtgtgcaac tccgggaacg ccgtttgttg ccgcctttgt acaaccccag
     3301 tcatcgtata taccggcatg tggaccgtta tacacaacgt agtagttgat atgagggtgt
     3361 tgaatacccg attctgctct gagaggagca actgtgctgt taagctcaga tttttgtggg
     3421 attggaattc ctccaagctt gaattcttgc agcgctccaa ttcagcaagc gtcgatctag
     3481 taacatagat gacaccgcgc gcgataattt atcctagttt gcgcgctata ttttgttttc
     3541 tatcgcgtat taaatgtata attgcgggac tctaatcata aaaacccatc tcataaataa
     3601 cgtcatgcat tacatgttaa ttattacatg cttaacgtaa ttcaacagaa attatatgat
     3661 aatcatcgca agaccggcaa caggattcaa tcttaagaaa ctttattgcc aaatgtttga
     3721 acgatctgct tgactctaga tccagagtcc cgctcaaatc tcagtcacag gaagaacagg
     3781 tctaggagga acaggaagag agaaatcaag ctgccagaat cccacatcat gccaatttcc
     3841 atgtttgaaa ccagcagctc taagcattcc cctaggagca taaccaagag cctcatgcat
     3901 cctcacagaa ggatcattag gaagtccaat cacagcaacc acagacttga aaccctgagc
     3961 ctccaaagac ttcaaaagat gagtgtaaag agtagatcca agaccagttc tttgatgcct
     4021 aggagacacg taaacagtag actcagcagt ccaatcgtaa gcattcctag ccttccaagg
     4081 accagcataa gcaataccag ccacttcacc atccacttca gcaacaagcc aaggatacct
     4141 ctctctaagc ctcacaagat catcagtcca ctcttgtggt tcttgaggct cagtcctgaa
     4201 attcacagta gaagtctcaa tgtaatgatt cacaatagtg cacacagcag gcatatcagc
     4261 ctcagtagcc cttctaatat cagcaggcct tctttcagga gacatgcgaa acgatccaga
     4321 tccggtgcag attatttgga ttgagagtga atatgagact ctaattggat accgagggga
     4381 atttatggaa cgtcagtgga gcattttgac aagaaatatt tgctagctga tagtgacctt
     4441 aggcgacttt tgaacgcgca ataatggttt ctgacgtatg tgcttagctc attaaactcc
     4501 agaaacccgc ggctgagtgg ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa
     4561 cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact cccttaattc tccgctccgg
     4621 atccgaattc ggcacagagt gttcaacccc agtacattaa aaacgtccgc aatgtgttat
     4681 taagttgtct aagcgtcaat ttgtttacac cacaatatat cctgccacca gccagccaac
     4741 agctccccga ccggcagctc ggcacaaaat caccactcga tacaggcagc ccatcaacca
     4801 tgactactgt ccggtccaac agatccgagc aggaaccgta aaaaggccgc gttgctggcg
     4861 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4921 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4981 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5041 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5101 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5161 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5221 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5281 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5341 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5401 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5461 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5521 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5581 aaatcaatct aaagtatata tgagtaaact tggtctgaca gctaaaacaa ttcatccagt
     5641 aaaatataat attttatttt ctcccaatca ggcttgatcc ccagtaagtc aaaaaatagc
     5701 tcgacatact gttcttcccc gatatcctcc ctgagcgacc ggacgcagaa ggcaatgtca
     5761 taccacttgt ccgccctgcc gcttctccca agatcaataa agccacttac tttgccatct
     5821 ttcacaaaga tgttgctgtc tcccaggtcg ccgtgggaaa agacaagttc ctcttcgggc
     5881 ttttccgtct ttaaaaaatc atacagctcg cgcggatctt taaatggagt gtcctcttcc
     5941 cagttttcgc aatccacatc ggccagatcg ttattcagta agtaatccaa ttcggctaag
     6001 cggctgtcta agctattcgt atagggacaa tccgatatgt cgatggagtg aaagagcctg
     6061 atgcactccg catacagctc gataatcttt tcagggcttt gttcatcttc atactcttcc
     6121 gagcaaagga cgccatcggc ctcactcatg agcagattgc tccagccatc atgccgttca
     6181 aagtgcagga cctttggaac aggcagcttt ccttccagcc atagcatcat gtccttttcc
     6241 cgttccacat cataggtggt ccctttatac cggctgtccg tcatttttaa atataggttt
     6301 tcattttctc ccaccagctt atatacctta gcaggagaca ttccttccgt atcttttacg
     6361 cagcggtatt tttcgatcag ttttttcaat tccggtgata ttctcatttt agccatactc
     6421 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6481 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcactcggag ccttgatgtg
     6541 ggcgcttttt gcagctgttc ggctgtgcgc tggccagaca gttatgcaca ggccaggcgg
     6601 gttttaagag ttttaataag ttttaaagag ttttaggcgg aaaaatcgcc ttttttctct
     6661 tttatatcag tcacttacat gtgtgaccgg ttcccaatgt acggctttgg gttcccaatg
     6721 tacgggttcc ggttcccaat gtacggcttt gggttcccaa tgtacgtgct atccacagga
     6781 aagaagcttt ttcgaccttt ttcccctgct agggcaattt gccctagcat ctgctccgta
     6841 cattaggaac cggcggatgc ttcgccctcg atcaggttgc ggtagcgcat gactaggatc
     6901 gggccagcct gccccgcctc ctccttcaaa tcgtactccg gcaggtcatt tgacccgatc
     6961 agcttgcgca cggtgaaaca gaacttcttg aactctccgg cgctgccact gcgttcgtag
     7021 atcgtcttga acaaccatct ggcttctgcc ttgcctgcgg cgcggcgtgc caggcggtag
     7081 agaaaacggc cgatgccggg atcgatcaaa aagtaatcgg ggtgaaccgt cagcacgtcc
     7141 gggttcttgc cttctgtgat ctcgcggtac atccaatcag ctagctcgat ctcgatgtac
     7201 tccggccgcc cggtttcgct ctttacgatc ttgtagcggc taatcaaggc ttcaccctcg
     7261 gataccgtca ccaggcggcc gttcttggcc ttcttcgtac gctgcatggc aacgtgcgtg
     7321 gtgtttaacc gaatgcaggt ttctaccagg tcgtctttct gctttccgcc atcggctcgc
     7381 cggcagaact tgagtacgtc cgcaacgtgt ggacggaaca cgcggccggg cttgtctccc
     7441 ttcccttccc ggtatcggtt catggattcg gttagatggg aaaccgccat cagtaccagg
     7501 tcgtaatccc acacactggc catgccggcc ggccctgcgg aaacctctac gtgcccgtct
     7561 ggaagctcgt agcggatcac ctcgccagct cgtcggtcac gcttcgacag acggaaaacg
     7621 gccacgtcca tgatgctgcg actatcgcgg gtgcccacgt catagagcat cggaacgaaa
     7681 aaatctggtt gctcgtcgcc cttgggcggc ttcctaatcg acgacgcacc ggctgccggc
     7741 ggttgccggg attctttgcg gattcgatca gcggccgctt gccacgattc accggggcgt
     7801 gcttctgcct cgatgcgttg ccgctgggcg gcctgcgcgg ccttcaactt ctccaccagg
     7861 tcatcaccca gcgccgcgcc gatttgtacc gggccggatg gtttgcgacc gctcacgccg
     7921 attcctcggg cttgggggtt ccagtgccat tgcagggccg gcagacaacc cagccgctta
     7981 cgcctggcca accgcccgtt cctccacaca tggggcattc cacggcgtcg gtgcctggtt
     8041 gttcttgatt ttccatgccg cctcctttag ccgctaaaat tcatctactc atttattcat
     8101 ttgctcattt actctggtag ctgcgcgatg tattcagata gcagctcggt aatggtcttg
     8161 ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct ccgccggcaa ctgaaagttg
     8221 acccgcttca tggctggcgt gtctgccagg ctggccaacg ttgcagcctt gctgctgcgt
     8281 gcgctcggac ggccggcact tagcgtgttt gtgcttttgc tcattttctc tttacctcat
     8341 taactcaaat gagttttgat ttaatttcag cggccagcgc ctggacctcg cgggcagcgt
     8401 cgccctcggg ttctgattca agaacggttg tgccggcggc ggcagtgcct gggtagctca
     8461 cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta cccggccagc gcctcggcaa
     8521 cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac gacaaaggcc gcttgtagcc
     8581 ttccatccgt gacctcaatg cgctgcttaa ccagctccac caggtcggcg gtggcccata
     8641 t
//
