LOCUS       sequence-342795-        8338 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 8338)
  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 8338)
  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_22804
FEATURES             Location/Qualifiers
     source          1..8338
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(247..876)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    1185..1209
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1276..1983)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2336..2681)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2373..2393)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3714..3966)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4566..4748)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4602..4780)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4943..4967
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5103..5691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5183..5202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5862..6656)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6657..6761)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6823..7017
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7083..8156)
                     /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"
ORIGIN
        1 atttgctcat ttactctggt agctgcgcga tgtattcaga tagcagctcg gtaatggtct
       61 tgccttggcg taccgcgtac atcttcagct tggtgtgatc ctccgccggc aactgaaagt
      121 tgacccgctt catggctggc gtgtctgcca ggctggccaa cgttgcagcc ttgctgctgc
      181 gtgcgctcgg acggccggca cttagcgtgt ttgtgctttt gctcattttc tctttacctc
      241 attaactcaa atgagttttg atttaatttc agcggccagc gcctggacct cgcgggcagc
      301 gtcgccctcg ggttctgatt caagaacggt tgtgccggcg gcggcagtgc ctgggtagct
      361 cacgcgctgc gtgatacggg actcaagaat gggcagctcg tacccggcca gcgcctcggc
      421 aacctcaccg ccgatgcgcg tgcctttgat cgcccgcgac acgacaaagg ccgcttgtag
      481 ccttccatcc gtgacctcaa tgcgctgctt aaccagctcc accaggtcgg cggtggccca
      541 tatgtcgtaa gggcttggct gcaccggaat cagcacgaag tcggctgcct tgatcgcgga
      601 cacagccaag tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc gccggccgat
      661 ggccttcacg tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc
      721 ttcccgcacg gccgcccaat cgcgggcact gccctgggga tcggaatcga ctaacagaac
      781 atcggccccg gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga
      841 cccgcctttc tggttaagta cagcgataac cttcatgcgt tccccttgcg tatttgttta
      901 tttactcatc gcatcatata cgcagcgacc gcatgacgca agctgtttta ctcaaataca
      961 catcaccttt ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc
     1021 gccagcttgg catcagacaa accggccagg atttcatgca gccgcacggc tgactcccga
     1081 ggggaaccct gtggttggca tgcacataca aatggacgaa cggataaacc ttttcacgcc
     1141 cttttaaata tccgattatt ctaataaacg ctcttttctc ttaggtttac ccgccaatat
     1201 atcctgtcaa acactgatag tttaaacgga tcacttcgtg tccctcggtc acatgtgcat
     1261 cctctagtag cggtcctgct gagcctcgac atgttgtcgc aaaattcgcc ctggacccgc
     1321 ccaacgattt gtcgtcactg tcaaggtttg acctgcactt catttggggc ccacatacac
     1381 caaaaaaatg ctgcataatt ctcggggcag caagtcggtt acccggccgc cgtgctggac
     1441 cgggttgaat ggtgcccgta actttcggta gagcggacgg ccaatactca acttcaagga
     1501 atctcaccca tgcgcgccgg cggggaaccg gagttccctt cagtgagcgt tattagttcg
     1561 ccgctcggtg tgtcgtagat actagcccct ggggcacttt tgaaatttga ataagattta
     1621 tgtaatcagt cttttaggtt tgaccggttc tgccgctttt tttaaaattg gatttgtaat
     1681 aataaaacgc aattgtttgt tattgtggcg ctctatcata gatgtcgcta taaacctatt
     1741 cagcacaata tattgttttc attttaatat tgtacatata agtagtaggg tacaatcagt
     1801 aaattgaacg gagaatatta ttcataaaaa tacgatagta acgggtgata tattcattag
     1861 aatgaaccga aaccggcggt aaggatctga gctacacatg ctcaggtttt ttacaacgtg
     1921 cacaacagaa ttgaaagcaa atatcatgcg atcataggct tctcgcatat ctcattaaag
     1981 caggacaagc tcacacagca aggtcagcgt ggttgttctt gatcaaccaa gatttgtaaa
     2041 actcagcgaa cctcttgaat ccagaaggag ttccccacct tccagagttc tgcatcctca
     2101 cagcagtgtc gtcaagcaca gctgagtaag tctttctctc cttcacaagg tcgttgaacc
     2161 taggctcaag cttagacatc tcccagttcc tgtacaacct cttcaacttc tcagtgatag
     2221 cagagaacct caaacccaca gacattgtat cgataattgt aaatgtaatt gtaatgttgt
     2281 ttgttgtttg ttgttgttgg taattgttgt aaaaatgagc tcttatactc gagcgtgtcc
     2341 tctccaaatg aaatgaactt ccttatatag aggaagggtc ttgcgaagga tagtgggatt
     2401 gtgcgtcatc ccttacgtca gtggagatgt cacatcaatc cacttgcttt gtagacgtgg
     2461 ttggaacctc ttctttttcc acgatgctcc tcgtgggtgg gggtccatct ttgggaccac
     2521 tgtcggcaga gagatcttga atgatagcct ttcctttatc gcaatgatgg catttgtagg
     2581 agccaccttc cttttctact gtcctttcga tgaagtgaca gatagctggg caatggaatc
     2641 cgaggaggtt tcccgaaatt atcctttgtt gaaaagtctc aatagccctt tgatcttctg
     2701 agactgtatc tttgacattt ttggagtaga ccagagtgtc gtgctccacc atgttgacga
     2761 agattttctt cttgtcattg agtcgtaaaa gactctgtat gaactgttcg ccagtcttga
     2821 cggcgagttc tgttagatcc tcgatttgaa tcttagactc catgcatggc cttagattca
     2881 gtaggaacta cctttttaga gactccaatc tctattactt gccttggttt atgaagcaag
     2941 ccttgaatcg tccatactgg aatagtactt ctgatcttga gaaatatgtc tttctctgtg
     3001 ttcttgatgc aattagtcct gaatcttttg actgcatctt taaccttctt gggaaggtat
     3061 ttgatctcct ggagattgtt actcgggtag atcgtcttga tgagacttgc tgcgtaggcc
     3121 tctctaacca tctgtgggtc agcattcttt ctgaaattga agaggctaac cttctcatta
     3181 tcagtggtga acatagtgtc gtcaccttca ccttcgaact tccttcctag atcgtaaaga
     3241 tagaggaaat cgtccattgt aatctccggg gcaaaggaga tctcttttgg ggctggatca
     3301 ctgctgggcc ttttggttcc tagcgtgagc cagtgggctt tttgctttgg tgggcttgtt
     3361 agggccttag caaagctctt gggcttgagt tgagcttctc ctttggggat gaagttcaac
     3421 ctgtctgttt gctgacttgt tgtgtacgcg tcagctgctg ctcttgcctc tgtaatagtg
     3481 gcaaatttct tgtgtgcaac tccgggaacg ccgtttgttg ccgcctttgt acaaccccag
     3541 tcatcgtata taccggcatg tggaccgtta tacacaacgt agtagttgat atgagggtgt
     3601 tgaatacccg attctgctct gagaggagca actgtgctgt taagctcaga tttttgtggg
     3661 attggaattc ctccaagctt gaattcttgc agcgctccaa ttcagcaagc gtcgatctag
     3721 taacatagat gacaccgcgc gcgataattt atcctagttt gcgcgctata ttttgttttc
     3781 tatcgcgtat taaatgtata attgcgggac tctaatcata aaaacccatc tcataaataa
     3841 cgtcatgcat tacatgttaa ttattacatg cttaacgtaa ttcaacagaa attatatgat
     3901 aatcatcgca agaccggcaa caggattcaa tcttaagaaa ctttattgcc aaatgtttga
     3961 acgatctgct tgactctaga tccagagtcc cgctcaaatc tcagtcacag gaagaacagg
     4021 tctaggagga acaggaagag agaaatcaag ctgccagaat cccacatcat gccaatttcc
     4081 atgtttgaaa ccagcagctc taagcattcc cctaggagca taaccaagag cctcatgcat
     4141 cctcacagaa ggatcattag gaagtccaat cacagcaacc acagacttga aaccctgagc
     4201 ctccaaagac ttcaaaagat gagtgtaaag agtagatcca agaccagttc tttgatgcct
     4261 aggagacacg taaacagtag actcagcagt ccaatcgtaa gcattcctag ccttccaagg
     4321 accagcataa gcaataccag ccacttcacc atccacttca gcaacaagcc aaggatacct
     4381 ctctctaagc ctcacaagat catcagtcca ctcttgtggt tcttgaggct cagtcctgaa
     4441 attcacagta gaagtctcaa tgtaatgatt cacaatagtg cacacagcag gcatatcagc
     4501 ctcagtagcc cttctaatat cagcaggcct tctttcagga gacatgcgaa acgatccaga
     4561 tccggtgcag attatttgga ttgagagtga atatgagact ctaattggat accgagggga
     4621 atttatggaa cgtcagtgga gcattttgac aagaaatatt tgctagctga tagtgacctt
     4681 aggcgacttt tgaacgcgca ataatggttt ctgacgtatg tgcttagctc attaaactcc
     4741 agaaacccgc ggctgagtgg ctccttcaac gttgcggttc tgtcagttcc aaacgtaaaa
     4801 cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact cccttaattc tccgctccgg
     4861 atccgaattc ggcacagagt gttcaacccc agtacattaa aaacgtccgc aatgtgttat
     4921 taagttgtct aagcgtcaat ttgtttacac cacaatatat cctgccacca gccagccaac
     4981 agctccccga ccggcagctc ggcacaaaat caccactcga tacaggcagc ccatcaacca
     5041 tgactactgt ccggtccaac agatccgagc aggaaccgta aaaaggccgc gttgctggcg
     5101 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5161 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5221 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5281 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5341 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5401 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5461 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5521 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5581 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5641 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5701 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5761 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5821 aaatcaatct aaagtatata tgagtaaact tggtctgaca gctaaaacaa ttcatccagt
     5881 aaaatataat attttatttt ctcccaatca ggcttgatcc ccagtaagtc aaaaaatagc
     5941 tcgacatact gttcttcccc gatatcctcc ctgagcgacc ggacgcagaa ggcaatgtca
     6001 taccacttgt ccgccctgcc gcttctccca agatcaataa agccacttac tttgccatct
     6061 ttcacaaaga tgttgctgtc tcccaggtcg ccgtgggaaa agacaagttc ctcttcgggc
     6121 ttttccgtct ttaaaaaatc atacagctcg cgcggatctt taaatggagt gtcctcttcc
     6181 cagttttcgc aatccacatc ggccagatcg ttattcagta agtaatccaa ttcggctaag
     6241 cggctgtcta agctattcgt atagggacaa tccgatatgt cgatggagtg aaagagcctg
     6301 atgcactccg catacagctc gataatcttt tcagggcttt gttcatcttc atactcttcc
     6361 gagcaaagga cgccatcggc ctcactcatg agcagattgc tccagccatc atgccgttca
     6421 aagtgcagga cctttggaac aggcagcttt ccttccagcc atagcatcat gtccttttcc
     6481 cgttccacat cataggtggt ccctttatac cggctgtccg tcatttttaa atataggttt
     6541 tcattttctc ccaccagctt atatacctta gcaggagaca ttccttccgt atcttttacg
     6601 cagcggtatt tttcgatcag ttttttcaat tccggtgata ttctcatttt agccatactc
     6661 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6721 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcactcggag ccttgatgtg
     6781 ggcgcttttt gcagctgttc ggctgtgcgc tggccagaca gttatgcaca ggccaggcgg
     6841 gttttaagag ttttaataag ttttaaagag ttttaggcgg aaaaatcgcc ttttttctct
     6901 tttatatcag tcacttacat gtgtgaccgg ttcccaatgt acggctttgg gttcccaatg
     6961 tacgggttcc ggttcccaat gtacggcttt gggttcccaa tgtacgtgct atccacagga
     7021 aagaagcttt ttcgaccttt ttcccctgct agggcaattt gccctagcat ctgctccgta
     7081 cattaggaac cggcggatgc ttcgccctcg atcaggttgc ggtagcgcat gactaggatc
     7141 gggccagcct gccccgcctc ctccttcaaa tcgtactccg gcaggtcatt tgacccgatc
     7201 agcttgcgca cggtgaaaca gaacttcttg aactctccgg cgctgccact gcgttcgtag
     7261 atcgtcttga acaaccatct ggcttctgcc ttgcctgcgg cgcggcgtgc caggcggtag
     7321 agaaaacggc cgatgccggg atcgatcaaa aagtaatcgg ggtgaaccgt cagcacgtcc
     7381 gggttcttgc cttctgtgat ctcgcggtac atccaatcag ctagctcgat ctcgatgtac
     7441 tccggccgcc cggtttcgct ctttacgatc ttgtagcggc taatcaaggc ttcaccctcg
     7501 gataccgtca ccaggcggcc gttcttggcc ttcttcgtac gctgcatggc aacgtgcgtg
     7561 gtgtttaacc gaatgcaggt ttctaccagg tcgtctttct gctttccgcc atcggctcgc
     7621 cggcagaact tgagtacgtc cgcaacgtgt ggacggaaca cgcggccggg cttgtctccc
     7681 ttcccttccc ggtatcggtt catggattcg gttagatggg aaaccgccat cagtaccagg
     7741 tcgtaatccc acacactggc catgccggcc ggccctgcgg aaacctctac gtgcccgtct
     7801 ggaagctcgt agcggatcac ctcgccagct cgtcggtcac gcttcgacag acggaaaacg
     7861 gccacgtcca tgatgctgcg actatcgcgg gtgcccacgt catagagcat cggaacgaaa
     7921 aaatctggtt gctcgtcgcc cttgggcggc ttcctaatcg acgacgcacc ggctgccggc
     7981 ggttgccggg attctttgcg gattcgatca gcggccgctt gccacgattc accggggcgt
     8041 gcttctgcct cgatgcgttg ccgctgggcg gcctgcgcgg ccttcaactt ctccaccagg
     8101 tcatcaccca gcgccgcgcc gatttgtacc gggccggatg gtttgcgacc gctcacgccg
     8161 attcctcggg cttgggggtt ccagtgccat tgcagggccg gcagacaacc cagccgctta
     8221 cgcctggcca accgcccgtt cctccacaca tggggcattc cacggcgtcg gtgcctggtt
     8281 gttcttgatt ttccatgccg cctcctttag ccgctaaaat tcatctactc atttattc
//
