LOCUS       sequence-342790-        8251 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 8251)
  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 8251)
  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_20301
FEATURES             Location/Qualifiers
     source          1..8251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(304..933)
                     /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    1242..1266
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1333..2040)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2306..2651)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2343..2363)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3684..3936)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4536..4718)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4572..4750)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4913..4937
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5073..5661)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5153..5172)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5832..6626)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6627..6731)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6793..6987
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7053..8126)
                     /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 ttcttgattt tccatgccgc ctcctttagc cgctaaaatt catctactca tttattcatt
       61 tgctcattta ctctggtagc tgcgcgatgt attcagatag cagctcggta atggtcttgc
      121 cttggcgtac cgcgtacatc ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga
      181 cccgcttcat ggctggcgtg tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg
      241 cgctcggacg gccggcactt agcgtgtttg tgcttttgct cattttctct ttacctcatt
      301 aactcaaatg agttttgatt taatttcagc ggccagcgcc tggacctcgc gggcagcgtc
      361 gccctcgggt tctgattcaa gaacggttgt gccggcggcg gcagtgcctg ggtagctcac
      421 gcgctgcgtg atacgggact caagaatggg cagctcgtac ccggccagcg cctcggcaac
      481 ctcaccgccg atgcgcgtgc ctttgatcgc ccgcgacacg acaaaggccg cttgtagcct
      541 tccatccgtg acctcaatgc gctgcttaac cagctccacc aggtcggcgg tggcccatat
      601 gtcgtaaggg cttggctgca ccggaatcag cacgaagtcg gctgccttga tcgcggacac
      661 agccaagtcc gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc
      721 cttcacgtcg cggtcaatcg tcgggcggtc gatgccgaca acggttagcg gttgatcttc
      781 ccgcacggcc gcccaatcgc gggcactgcc ctggggatcg gaatcgacta acagaacatc
      841 ggccccggcg agttgcaggg cgcgggctag atgggttgcg atggtcgtct tgcctgaccc
      901 gcctttctgg ttaagtacag cgataacctt catgcgttcc ccttgcgtat ttgtttattt
      961 actcatcgca tcatatacgc agcgaccgca tgacgcaagc tgttttactc aaatacacat
     1021 caccttttta gacggcggcg ctcggtttct tcagcggcca agctggccgg ccaggccgcc
     1081 agcttggcat cagacaaacc ggccaggatt tcatgcagcc gcacggctga ctcccgaggg
     1141 gaaccctgtg gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt
     1201 ttaaatatcc gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc
     1261 ctgtcaaaca ctgatagttt aaacggatca cttcgtgtcc ctcggtcaca tgtgcatcct
     1321 ctagtagcgg tcctgctgag cctcgacatg ttgtcgcaaa attcgccctg gacccgccca
     1381 acgatttgtc gtcactgtca aggtttgacc tgcacttcat ttggggccca catacaccaa
     1441 aaaaatgctg cataattctc ggggcagcaa gtcggttacc cggccgccgt gctggaccgg
     1501 gttgaatggt gcccgtaact ttcggtagag cggacggcca atactcaact tcaaggaatc
     1561 tcacccatgc gcgccggcgg ggaaccggag ttcccttcag tgagcgttat tagttcgccg
     1621 ctcggtgtgt cgtagatact agcccctggg gcacttttga aatttgaata agatttatgt
     1681 aatcagtctt ttaggtttga ccggttctgc cgcttttttt aaaattggat ttgtaataat
     1741 aaaacgcaat tgtttgttat tgtggcgctc tatcatagat gtcgctataa acctattcag
     1801 cacaatatat tgttttcatt ttaatattgt acatataagt agtagggtac aatcagtaaa
     1861 ttgaacggag aatattattc ataaaaatac gatagtaacg ggtgatatat tcattagaat
     1921 gaaccgaaac cggcggtaag gatctgagct acacatgctc aggtttttta caacgtgcac
     1981 aacagaattg aaagcaaata tcatgcgatc ataggcttct cgcatatctc attaaagcag
     2041 gacaagctca agacttagtc atcttagcct tagccttctt agcagcagcg atcaacttag
     2101 agatcttgat gtcaggcttc ttgatgatct tgttagtaga agggaacacg ttcttccaga
     2161 atcctccgta tccgtcaccc cttccagact ggatcacctc ctgagcctcc accattgtat
     2221 cgataattgt aaatgtaatt gtaatgttgt ttgttgtttg ttgttgttgg taattgttgt
     2281 aaaaatgagc tcttatactc gagcgtgtcc tctccaaatg aaatgaactt ccttatatag
     2341 aggaagggtc ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca gtggagatgt
     2401 cacatcaatc cacttgcttt gtagacgtgg ttggaacctc ttctttttcc acgatgctcc
     2461 tcgtgggtgg gggtccatct ttgggaccac tgtcggcaga gagatcttga atgatagcct
     2521 ttcctttatc gcaatgatgg catttgtagg agccaccttc cttttctact gtcctttcga
     2581 tgaagtgaca gatagctggg caatggaatc cgaggaggtt tcccgaaatt atcctttgtt
     2641 gaaaagtctc aatagccctt tgatcttctg agactgtatc tttgacattt ttggagtaga
     2701 ccagagtgtc gtgctccacc atgttgacga agattttctt cttgtcattg agtcgtaaaa
     2761 gactctgtat gaactgttcg ccagtcttga cggcgagttc tgttagatcc tcgatttgaa
     2821 tcttagactc catgcatggc cttagattca gtaggaacta cctttttaga gactccaatc
     2881 tctattactt gccttggttt atgaagcaag ccttgaatcg tccatactgg aatagtactt
     2941 ctgatcttga gaaatatgtc tttctctgtg ttcttgatgc aattagtcct gaatcttttg
     3001 actgcatctt taaccttctt gggaaggtat ttgatctcct ggagattgtt actcgggtag
     3061 atcgtcttga tgagacttgc tgcgtaggcc tctctaacca tctgtgggtc agcattcttt
     3121 ctgaaattga agaggctaac cttctcatta tcagtggtga acatagtgtc gtcaccttca
     3181 ccttcgaact tccttcctag atcgtaaaga tagaggaaat cgtccattgt aatctccggg
     3241 gcaaaggaga tctcttttgg ggctggatca ctgctgggcc ttttggttcc tagcgtgagc
     3301 cagtgggctt tttgctttgg tgggcttgtt agggccttag caaagctctt gggcttgagt
     3361 tgagcttctc ctttggggat gaagttcaac ctgtctgttt gctgacttgt tgtgtacgcg
     3421 tcagctgctg ctcttgcctc tgtaatagtg gcaaatttct tgtgtgcaac tccgggaacg
     3481 ccgtttgttg ccgcctttgt acaaccccag tcatcgtata taccggcatg tggaccgtta
     3541 tacacaacgt agtagttgat atgagggtgt tgaatacccg attctgctct gagaggagca
     3601 actgtgctgt taagctcaga tttttgtggg attggaattc ctccaagctt gaattcttgc
     3661 agcgctccaa ttcagcaagc gtcgatctag taacatagat gacaccgcgc gcgataattt
     3721 atcctagttt gcgcgctata ttttgttttc tatcgcgtat taaatgtata attgcgggac
     3781 tctaatcata aaaacccatc tcataaataa cgtcatgcat tacatgttaa ttattacatg
     3841 cttaacgtaa ttcaacagaa attatatgat aatcatcgca agaccggcaa caggattcaa
     3901 tcttaagaaa ctttattgcc aaatgtttga acgatctgct tgactctaga tccagagtcc
     3961 cgctcaaatc tcagtcacag gaagaacagg tctaggagga acaggaagag agaaatcaag
     4021 ctgccagaat cccacatcat gccaatttcc atgtttgaaa ccagcagctc taagcattcc
     4081 cctaggagca taaccaagag cctcatgcat cctcacagaa ggatcattag gaagtccaat
     4141 cacagcaacc acagacttga aaccctgagc ctccaaagac ttcaaaagat gagtgtaaag
     4201 agtagatcca agaccagttc tttgatgcct aggagacacg taaacagtag actcagcagt
     4261 ccaatcgtaa gcattcctag ccttccaagg accagcataa gcaataccag ccacttcacc
     4321 atccacttca gcaacaagcc aaggatacct ctctctaagc ctcacaagat catcagtcca
     4381 ctcttgtggt tcttgaggct cagtcctgaa attcacagta gaagtctcaa tgtaatgatt
     4441 cacaatagtg cacacagcag gcatatcagc ctcagtagcc cttctaatat cagcaggcct
     4501 tctttcagga gacatgcgaa acgatccaga tccggtgcag attatttgga ttgagagtga
     4561 atatgagact ctaattggat accgagggga atttatggaa cgtcagtgga gcattttgac
     4621 aagaaatatt tgctagctga tagtgacctt aggcgacttt tgaacgcgca ataatggttt
     4681 ctgacgtatg tgcttagctc attaaactcc agaaacccgc ggctgagtgg ctccttcaac
     4741 gttgcggttc tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca
     4801 taacgtgact cccttaattc tccgctccgg atccgaattc ggcacagagt gttcaacccc
     4861 agtacattaa aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac
     4921 cacaatatat cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat
     4981 caccactcga tacaggcagc ccatcaacca tgactactgt ccggtccaac agatccgagc
     5041 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5101 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5161 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5221 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5281 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5341 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5401 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5461 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5521 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5581 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5641 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5701 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5761 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5821 tggtctgaca gctaaaacaa ttcatccagt aaaatataat attttatttt ctcccaatca
     5881 ggcttgatcc ccagtaagtc aaaaaatagc tcgacatact gttcttcccc gatatcctcc
     5941 ctgagcgacc ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc gcttctccca
     6001 agatcaataa agccacttac tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg
     6061 ccgtgggaaa agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg
     6121 cgcggatctt taaatggagt gtcctcttcc cagttttcgc aatccacatc ggccagatcg
     6181 ttattcagta agtaatccaa ttcggctaag cggctgtcta agctattcgt atagggacaa
     6241 tccgatatgt cgatggagtg aaagagcctg atgcactccg catacagctc gataatcttt
     6301 tcagggcttt gttcatcttc atactcttcc gagcaaagga cgccatcggc ctcactcatg
     6361 agcagattgc tccagccatc atgccgttca aagtgcagga cctttggaac aggcagcttt
     6421 ccttccagcc atagcatcat gtccttttcc cgttccacat cataggtggt ccctttatac
     6481 cggctgtccg tcatttttaa atataggttt tcattttctc ccaccagctt atatacctta
     6541 gcaggagaca ttccttccgt atcttttacg cagcggtatt tttcgatcag ttttttcaat
     6601 tccggtgata ttctcatttt agccatactc ttcctttttc aatattattg aagcatttat
     6661 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6721 ggggttccgc gcactcggag ccttgatgtg ggcgcttttt gcagctgttc ggctgtgcgc
     6781 tggccagaca gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag
     6841 ttttaggcgg aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg
     6901 ttcccaatgt acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt
     6961 gggttcccaa tgtacgtgct atccacagga aagaagcttt ttcgaccttt ttcccctgct
     7021 agggcaattt gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg
     7081 atcaggttgc ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa
     7141 tcgtactccg gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg
     7201 aactctccgg cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc
     7261 ttgcctgcgg cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa
     7321 aagtaatcgg ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat ctcgcggtac
     7381 atccaatcag ctagctcgat ctcgatgtac tccggccgcc cggtttcgct ctttacgatc
     7441 ttgtagcggc taatcaaggc ttcaccctcg gataccgtca ccaggcggcc gttcttggcc
     7501 ttcttcgtac gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt ttctaccagg
     7561 tcgtctttct gctttccgcc atcggctcgc cggcagaact tgagtacgtc cgcaacgtgt
     7621 ggacggaaca cgcggccggg cttgtctccc ttcccttccc ggtatcggtt catggattcg
     7681 gttagatggg aaaccgccat cagtaccagg tcgtaatccc acacactggc catgccggcc
     7741 ggccctgcgg aaacctctac gtgcccgtct ggaagctcgt agcggatcac ctcgccagct
     7801 cgtcggtcac gcttcgacag acggaaaacg gccacgtcca tgatgctgcg actatcgcgg
     7861 gtgcccacgt catagagcat cggaacgaaa aaatctggtt gctcgtcgcc cttgggcggc
     7921 ttcctaatcg acgacgcacc ggctgccggc ggttgccggg attctttgcg gattcgatca
     7981 gcggccgctt gccacgattc accggggcgt gcttctgcct cgatgcgttg ccgctgggcg
     8041 gcctgcgcgg ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc gatttgtacc
     8101 gggccggatg gtttgcgacc gctcacgccg attcctcggg cttgggggtt ccagtgccat
     8161 tgcagggccg gcagacaacc cagccgctta cgcctggcca accgcccgtt cctccacaca
     8221 tggggcattc cacggcgtcg gtgcctggtt g
//
