LOCUS       sequence-342752-        8389 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 8389)
  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 8389)
  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_16275
FEATURES             Location/Qualifiers
     source          1..8389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    634..658
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(725..1432)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1836..2181)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1873..1893)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3214..3466)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4066..4248)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4102..4280)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4443..4467
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4603..5191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4683..4702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5362..6156)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6157..6261)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6323..6517
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6583..7656)
                     /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(8085..8389,1..325))
                     /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 ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt
       61 ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt
      121 cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg
      181 ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg
      241 cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct
      301 ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg
      361 catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt
      421 tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc
      481 atcagacaaa ccggccagga tttcatgcag ccgcacggct gactcccgag gggaaccctg
      541 tggttggcat gcacatacaa atggacgaac ggataaacct tttcacgccc ttttaaatat
      601 ccgattattc taataaacgc tcttttctct taggtttacc cgccaatata tcctgtcaaa
      661 cactgatagt ttaaacggat cacttcgtgt ccctcggtca catgtgcatc ctctagtagc
      721 ggtcctgctg agcctcgaca tgttgtcgca aaattcgccc tggacccgcc caacgatttg
      781 tcgtcactgt caaggtttga cctgcacttc atttggggcc cacatacacc aaaaaaatgc
      841 tgcataattc tcggggcagc aagtcggtta cccggccgcc gtgctggacc gggttgaatg
      901 gtgcccgtaa ctttcggtag agcggacggc caatactcaa cttcaaggaa tctcacccat
      961 gcgcgccggc ggggaaccgg agttcccttc agtgagcgtt attagttcgc cgctcggtgt
     1021 gtcgtagata ctagcccctg gggcactttt gaaatttgaa taagatttat gtaatcagtc
     1081 ttttaggttt gaccggttct gccgcttttt ttaaaattgg atttgtaata ataaaacgca
     1141 attgtttgtt attgtggcgc tctatcatag atgtcgctat aaacctattc agcacaatat
     1201 attgttttca ttttaatatt gtacatataa gtagtagggt acaatcagta aattgaacgg
     1261 agaatattat tcataaaaat acgatagtaa cgggtgatat attcattaga atgaaccgaa
     1321 accggcggta aggatctgag ctacacatgc tcaggttttt tacaacgtgc acaacagaat
     1381 tgaaagcaaa tatcatgcga tcataggctt ctcgcatatc tcattaaagc aggacaagct
     1441 cagtccaact ttcccattgt tttaaggtaa ttagtgtact caagcactgt tttcatcttc
     1501 tcagcgtcag tgaagtcagg gaagtccttc acgatcttag ccatgtcagc agggttcatc
     1561 ttagcctcct cagcgaactt gaaagctccc ttcatcttgt cgtcagccaa cacaagagcc
     1621 ttctccaaca caggcaagtt caatgttttt tccaacttct ccaaagctga gaaagagtta
     1681 gtcctgtaca ccttgtccac cacgaactta gccattgcct cctcagccac ctccttgaat
     1741 cccattgtat cgataattgt aaatgtaatt gtaatgttgt ttgttgtttg ttgttgttgg
     1801 taattgttgt aaaaatgagc tcttatactc gagcgtgtcc tctccaaatg aaatgaactt
     1861 ccttatatag aggaagggtc ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca
     1921 gtggagatgt cacatcaatc cacttgcttt gtagacgtgg ttggaacctc ttctttttcc
     1981 acgatgctcc tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga gagatcttga
     2041 atgatagcct ttcctttatc gcaatgatgg catttgtagg agccaccttc cttttctact
     2101 gtcctttcga tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgaaatt
     2161 atcctttgtt gaaaagtctc aatagccctt tgatcttctg agactgtatc tttgacattt
     2221 ttggagtaga ccagagtgtc gtgctccacc atgttgacga agattttctt cttgtcattg
     2281 agtcgtaaaa gactctgtat gaactgttcg ccagtcttga cggcgagttc tgttagatcc
     2341 tcgatttgaa tcttagactc catgcatggc cttagattca gtaggaacta cctttttaga
     2401 gactccaatc tctattactt gccttggttt atgaagcaag ccttgaatcg tccatactgg
     2461 aatagtactt ctgatcttga gaaatatgtc tttctctgtg ttcttgatgc aattagtcct
     2521 gaatcttttg actgcatctt taaccttctt gggaaggtat ttgatctcct ggagattgtt
     2581 actcgggtag atcgtcttga tgagacttgc tgcgtaggcc tctctaacca tctgtgggtc
     2641 agcattcttt ctgaaattga agaggctaac cttctcatta tcagtggtga acatagtgtc
     2701 gtcaccttca ccttcgaact tccttcctag atcgtaaaga tagaggaaat cgtccattgt
     2761 aatctccggg gcaaaggaga tctcttttgg ggctggatca ctgctgggcc ttttggttcc
     2821 tagcgtgagc cagtgggctt tttgctttgg tgggcttgtt agggccttag caaagctctt
     2881 gggcttgagt tgagcttctc ctttggggat gaagttcaac ctgtctgttt gctgacttgt
     2941 tgtgtacgcg tcagctgctg ctcttgcctc tgtaatagtg gcaaatttct tgtgtgcaac
     3001 tccgggaacg ccgtttgttg ccgcctttgt acaaccccag tcatcgtata taccggcatg
     3061 tggaccgtta tacacaacgt agtagttgat atgagggtgt tgaatacccg attctgctct
     3121 gagaggagca actgtgctgt taagctcaga tttttgtggg attggaattc ctccaagctt
     3181 gaattcttgc agcgctccaa ttcagcaagc gtcgatctag taacatagat gacaccgcgc
     3241 gcgataattt atcctagttt gcgcgctata ttttgttttc tatcgcgtat taaatgtata
     3301 attgcgggac tctaatcata aaaacccatc tcataaataa cgtcatgcat tacatgttaa
     3361 ttattacatg cttaacgtaa ttcaacagaa attatatgat aatcatcgca agaccggcaa
     3421 caggattcaa tcttaagaaa ctttattgcc aaatgtttga acgatctgct tgactctaga
     3481 tccagagtcc cgctcaaatc tcagtcacag gaagaacagg tctaggagga acaggaagag
     3541 agaaatcaag ctgccagaat cccacatcat gccaatttcc atgtttgaaa ccagcagctc
     3601 taagcattcc cctaggagca taaccaagag cctcatgcat cctcacagaa ggatcattag
     3661 gaagtccaat cacagcaacc acagacttga aaccctgagc ctccaaagac ttcaaaagat
     3721 gagtgtaaag agtagatcca agaccagttc tttgatgcct aggagacacg taaacagtag
     3781 actcagcagt ccaatcgtaa gcattcctag ccttccaagg accagcataa gcaataccag
     3841 ccacttcacc atccacttca gcaacaagcc aaggatacct ctctctaagc ctcacaagat
     3901 catcagtcca ctcttgtggt tcttgaggct cagtcctgaa attcacagta gaagtctcaa
     3961 tgtaatgatt cacaatagtg cacacagcag gcatatcagc ctcagtagcc cttctaatat
     4021 cagcaggcct tctttcagga gacatgcgaa acgatccaga tccggtgcag attatttgga
     4081 ttgagagtga atatgagact ctaattggat accgagggga atttatggaa cgtcagtgga
     4141 gcattttgac aagaaatatt tgctagctga tagtgacctt aggcgacttt tgaacgcgca
     4201 ataatggttt ctgacgtatg tgcttagctc attaaactcc agaaacccgc ggctgagtgg
     4261 ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg
     4321 gcgggggtca taacgtgact cccttaattc tccgctccgg atccgaattc ggcacagagt
     4381 gttcaacccc agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat
     4441 ttgtttacac cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc
     4501 ggcacaaaat caccactcga tacaggcagc ccatcaacca tgactactgt ccggtccaac
     4561 agatccgagc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4621 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4681 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4741 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4801 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4861 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4921 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4981 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5041 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5101 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5161 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5221 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5281 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5341 tgagtaaact tggtctgaca gctaaaacaa ttcatccagt aaaatataat attttatttt
     5401 ctcccaatca ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc
     5461 gatatcctcc ctgagcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc
     5521 gcttctccca agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc
     5581 tcccaggtcg ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc
     5641 atacagctcg cgcggatctt taaatggagt gtcctcttcc cagttttcgc aatccacatc
     5701 ggccagatcg ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt
     5761 atagggacaa tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc
     5821 gataatcttt tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc
     5881 ctcactcatg agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac
     5941 aggcagcttt ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt
     6001 ccctttatac cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt
     6061 atatacctta gcaggagaca ttccttccgt atcttttacg cagcggtatt tttcgatcag
     6121 ttttttcaat tccggtgata ttctcatttt agccatactc ttcctttttc aatattattg
     6181 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6241 taaacaaata ggggttccgc gcactcggag ccttgatgtg ggcgcttttt gcagctgttc
     6301 ggctgtgcgc tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag
     6361 ttttaaagag ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat
     6421 gtgtgaccgg ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat
     6481 gtacggcttt gggttcccaa tgtacgtgct atccacagga aagaagcttt ttcgaccttt
     6541 ttcccctgct agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc
     6601 ttcgccctcg atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc
     6661 ctccttcaaa tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca
     6721 gaacttcttg aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct
     6781 ggcttctgcc ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg
     6841 atcgatcaaa aagtaatcgg ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat
     6901 ctcgcggtac atccaatcag ctagctcgat ctcgatgtac tccggccgcc cggtttcgct
     6961 ctttacgatc ttgtagcggc taatcaaggc ttcaccctcg gataccgtca ccaggcggcc
     7021 gttcttggcc ttcttcgtac gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt
     7081 ttctaccagg tcgtctttct gctttccgcc atcggctcgc cggcagaact tgagtacgtc
     7141 cgcaacgtgt ggacggaaca cgcggccggg cttgtctccc ttcccttccc ggtatcggtt
     7201 catggattcg gttagatggg aaaccgccat cagtaccagg tcgtaatccc acacactggc
     7261 catgccggcc ggccctgcgg aaacctctac gtgcccgtct ggaagctcgt agcggatcac
     7321 ctcgccagct cgtcggtcac gcttcgacag acggaaaacg gccacgtcca tgatgctgcg
     7381 actatcgcgg gtgcccacgt catagagcat cggaacgaaa aaatctggtt gctcgtcgcc
     7441 cttgggcggc ttcctaatcg acgacgcacc ggctgccggc ggttgccggg attctttgcg
     7501 gattcgatca gcggccgctt gccacgattc accggggcgt gcttctgcct cgatgcgttg
     7561 ccgctgggcg gcctgcgcgg ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc
     7621 gatttgtacc gggccggatg gtttgcgacc gctcacgccg attcctcggg cttgggggtt
     7681 ccagtgccat tgcagggccg gcagacaacc cagccgctta cgcctggcca accgcccgtt
     7741 cctccacaca tggggcattc cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg
     7801 cctcctttag ccgctaaaat tcatctactc atttattcat ttgctcattt actctggtag
     7861 ctgcgcgatg tattcagata gcagctcggt aatggtcttg ccttggcgta ccgcgtacat
     7921 cttcagcttg gtgtgatcct ccgccggcaa ctgaaagttg acccgcttca tggctggcgt
     7981 gtctgccagg ctggccaacg ttgcagcctt gctgctgcgt gcgctcggac ggccggcact
     8041 tagcgtgttt gtgcttttgc tcattttctc tttacctcat taactcaaat gagttttgat
     8101 ttaatttcag cggccagcgc ctggacctcg cgggcagcgt cgccctcggg ttctgattca
     8161 agaacggttg tgccggcggc ggcagtgcct gggtagctca cgcgctgcgt gatacgggac
     8221 tcaagaatgg gcagctcgta cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg
     8281 cctttgatcg cccgcgacac gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg
     8341 cgctgcttaa ccagctccac caggtcggcg gtggcccata tgtcgtaag
//
