LOCUS       sequence-342793-        8329 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 8329)
  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 8329)
  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_21388
FEATURES             Location/Qualifiers
     source          1..8329
                     /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(1776..2121)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1813..1833)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3154..3406)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4006..4188)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4042..4220)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4383..4407
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4543..5131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4623..4642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5302..6096)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6097..6201)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6263..6457
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6523..7596)
                     /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(8025..8329,1..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 caagaaggac ccacgttctt gtcgtaccac ttcttgaacc tgttgaagaa cacagaagca
     1501 gaagccttgt caaggtgctt agtcctaaga gatctaggag tctctccaga cttgtacaaa
     1561 gcagtgaaaa gcttgtcctc gtacctaagc acagtctttg actgagtcct aggagagaat
     1621 cccaacttga atccgtaaag caccatcttc acgtactcag cagacttaga gatagagaaa
     1681 gccattgtat cgataattgt aaatgtaatt gtaatgttgt ttgttgtttg ttgttgttgg
     1741 taattgttgt aaaaatgagc tcttatactc gagcgtgtcc tctccaaatg aaatgaactt
     1801 ccttatatag aggaagggtc ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca
     1861 gtggagatgt cacatcaatc cacttgcttt gtagacgtgg ttggaacctc ttctttttcc
     1921 acgatgctcc tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga gagatcttga
     1981 atgatagcct ttcctttatc gcaatgatgg catttgtagg agccaccttc cttttctact
     2041 gtcctttcga tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgaaatt
     2101 atcctttgtt gaaaagtctc aatagccctt tgatcttctg agactgtatc tttgacattt
     2161 ttggagtaga ccagagtgtc gtgctccacc atgttgacga agattttctt cttgtcattg
     2221 agtcgtaaaa gactctgtat gaactgttcg ccagtcttga cggcgagttc tgttagatcc
     2281 tcgatttgaa tcttagactc catgcatggc cttagattca gtaggaacta cctttttaga
     2341 gactccaatc tctattactt gccttggttt atgaagcaag ccttgaatcg tccatactgg
     2401 aatagtactt ctgatcttga gaaatatgtc tttctctgtg ttcttgatgc aattagtcct
     2461 gaatcttttg actgcatctt taaccttctt gggaaggtat ttgatctcct ggagattgtt
     2521 actcgggtag atcgtcttga tgagacttgc tgcgtaggcc tctctaacca tctgtgggtc
     2581 agcattcttt ctgaaattga agaggctaac cttctcatta tcagtggtga acatagtgtc
     2641 gtcaccttca ccttcgaact tccttcctag atcgtaaaga tagaggaaat cgtccattgt
     2701 aatctccggg gcaaaggaga tctcttttgg ggctggatca ctgctgggcc ttttggttcc
     2761 tagcgtgagc cagtgggctt tttgctttgg tgggcttgtt agggccttag caaagctctt
     2821 gggcttgagt tgagcttctc ctttggggat gaagttcaac ctgtctgttt gctgacttgt
     2881 tgtgtacgcg tcagctgctg ctcttgcctc tgtaatagtg gcaaatttct tgtgtgcaac
     2941 tccgggaacg ccgtttgttg ccgcctttgt acaaccccag tcatcgtata taccggcatg
     3001 tggaccgtta tacacaacgt agtagttgat atgagggtgt tgaatacccg attctgctct
     3061 gagaggagca actgtgctgt taagctcaga tttttgtggg attggaattc ctccaagctt
     3121 gaattcttgc agcgctccaa ttcagcaagc gtcgatctag taacatagat gacaccgcgc
     3181 gcgataattt atcctagttt gcgcgctata ttttgttttc tatcgcgtat taaatgtata
     3241 attgcgggac tctaatcata aaaacccatc tcataaataa cgtcatgcat tacatgttaa
     3301 ttattacatg cttaacgtaa ttcaacagaa attatatgat aatcatcgca agaccggcaa
     3361 caggattcaa tcttaagaaa ctttattgcc aaatgtttga acgatctgct tgactctaga
     3421 tccagagtcc cgctcaaatc tcagtcacag gaagaacagg tctaggagga acaggaagag
     3481 agaaatcaag ctgccagaat cccacatcat gccaatttcc atgtttgaaa ccagcagctc
     3541 taagcattcc cctaggagca taaccaagag cctcatgcat cctcacagaa ggatcattag
     3601 gaagtccaat cacagcaacc acagacttga aaccctgagc ctccaaagac ttcaaaagat
     3661 gagtgtaaag agtagatcca agaccagttc tttgatgcct aggagacacg taaacagtag
     3721 actcagcagt ccaatcgtaa gcattcctag ccttccaagg accagcataa gcaataccag
     3781 ccacttcacc atccacttca gcaacaagcc aaggatacct ctctctaagc ctcacaagat
     3841 catcagtcca ctcttgtggt tcttgaggct cagtcctgaa attcacagta gaagtctcaa
     3901 tgtaatgatt cacaatagtg cacacagcag gcatatcagc ctcagtagcc cttctaatat
     3961 cagcaggcct tctttcagga gacatgcgaa acgatccaga tccggtgcag attatttgga
     4021 ttgagagtga atatgagact ctaattggat accgagggga atttatggaa cgtcagtgga
     4081 gcattttgac aagaaatatt tgctagctga tagtgacctt aggcgacttt tgaacgcgca
     4141 ataatggttt ctgacgtatg tgcttagctc attaaactcc agaaacccgc ggctgagtgg
     4201 ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg
     4261 gcgggggtca taacgtgact cccttaattc tccgctccgg atccgaattc ggcacagagt
     4321 gttcaacccc agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat
     4381 ttgtttacac cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc
     4441 ggcacaaaat caccactcga tacaggcagc ccatcaacca tgactactgt ccggtccaac
     4501 agatccgagc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4561 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4621 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4681 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4741 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4801 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4861 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4921 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4981 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5041 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5101 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5161 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5221 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5281 tgagtaaact tggtctgaca gctaaaacaa ttcatccagt aaaatataat attttatttt
     5341 ctcccaatca ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc
     5401 gatatcctcc ctgagcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc
     5461 gcttctccca agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc
     5521 tcccaggtcg ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc
     5581 atacagctcg cgcggatctt taaatggagt gtcctcttcc cagttttcgc aatccacatc
     5641 ggccagatcg ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt
     5701 atagggacaa tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc
     5761 gataatcttt tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc
     5821 ctcactcatg agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac
     5881 aggcagcttt ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt
     5941 ccctttatac cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt
     6001 atatacctta gcaggagaca ttccttccgt atcttttacg cagcggtatt tttcgatcag
     6061 ttttttcaat tccggtgata ttctcatttt agccatactc ttcctttttc aatattattg
     6121 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6181 taaacaaata ggggttccgc gcactcggag ccttgatgtg ggcgcttttt gcagctgttc
     6241 ggctgtgcgc tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag
     6301 ttttaaagag ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat
     6361 gtgtgaccgg ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat
     6421 gtacggcttt gggttcccaa tgtacgtgct atccacagga aagaagcttt ttcgaccttt
     6481 ttcccctgct agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc
     6541 ttcgccctcg atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc
     6601 ctccttcaaa tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca
     6661 gaacttcttg aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct
     6721 ggcttctgcc ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg
     6781 atcgatcaaa aagtaatcgg ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat
     6841 ctcgcggtac atccaatcag ctagctcgat ctcgatgtac tccggccgcc cggtttcgct
     6901 ctttacgatc ttgtagcggc taatcaaggc ttcaccctcg gataccgtca ccaggcggcc
     6961 gttcttggcc ttcttcgtac gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt
     7021 ttctaccagg tcgtctttct gctttccgcc atcggctcgc cggcagaact tgagtacgtc
     7081 cgcaacgtgt ggacggaaca cgcggccggg cttgtctccc ttcccttccc ggtatcggtt
     7141 catggattcg gttagatggg aaaccgccat cagtaccagg tcgtaatccc acacactggc
     7201 catgccggcc ggccctgcgg aaacctctac gtgcccgtct ggaagctcgt agcggatcac
     7261 ctcgccagct cgtcggtcac gcttcgacag acggaaaacg gccacgtcca tgatgctgcg
     7321 actatcgcgg gtgcccacgt catagagcat cggaacgaaa aaatctggtt gctcgtcgcc
     7381 cttgggcggc ttcctaatcg acgacgcacc ggctgccggc ggttgccggg attctttgcg
     7441 gattcgatca gcggccgctt gccacgattc accggggcgt gcttctgcct cgatgcgttg
     7501 ccgctgggcg gcctgcgcgg ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc
     7561 gatttgtacc gggccggatg gtttgcgacc gctcacgccg attcctcggg cttgggggtt
     7621 ccagtgccat tgcagggccg gcagacaacc cagccgctta cgcctggcca accgcccgtt
     7681 cctccacaca tggggcattc cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg
     7741 cctcctttag ccgctaaaat tcatctactc atttattcat ttgctcattt actctggtag
     7801 ctgcgcgatg tattcagata gcagctcggt aatggtcttg ccttggcgta ccgcgtacat
     7861 cttcagcttg gtgtgatcct ccgccggcaa ctgaaagttg acccgcttca tggctggcgt
     7921 gtctgccagg ctggccaacg ttgcagcctt gctgctgcgt gcgctcggac ggccggcact
     7981 tagcgtgttt gtgcttttgc tcattttctc tttacctcat taactcaaat gagttttgat
     8041 ttaatttcag cggccagcgc ctggacctcg cgggcagcgt cgccctcggg ttctgattca
     8101 agaacggttg tgccggcggc ggcagtgcct gggtagctca cgcgctgcgt gatacgggac
     8161 tcaagaatgg gcagctcgta cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg
     8221 cctttgatcg cccgcgacac gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg
     8281 cgctgcttaa ccagctccac caggtcggcg gtggcccata tgtcgtaag
//
