LOCUS       sequence-342337-        8692 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 8692)
  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 8692)
  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_13048
FEATURES             Location/Qualifiers
     source          1..8692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    671..695
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(762..1469)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2176..2521)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2213..2233)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3554..3806)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4406..4588)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4442..4620)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4783..4807
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4943..5531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5023..5042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5702..6496)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6497..6601)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6663..6857
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6923..7996)
                     /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(8425..8692,1..362))
                     /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 agctccacca ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac cggaatcagc
       61 acgaagtcgg ctgccttgat cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg
      121 atcactacga agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg
      181 atgccgacaa cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc
      241 tggggatcgg aatcgactaa cagaacatcg gccccggcga gttgcagggc gcgggctaga
      301 tgggttgcga tggtcgtctt gcctgacccg cctttctggt taagtacagc gataaccttc
      361 atgcgttccc cttgcgtatt tgtttattta ctcatcgcat catatacgca gcgaccgcat
      421 gacgcaagct gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt
      481 cagcggccaa gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt
      541 catgcagccg cacggctgac tcccgagggg aaccctgtgg ttggcatgca catacaaatg
      601 gacgaacgga taaacctttt cacgcccttt taaatatccg attattctaa taaacgctct
      661 tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta aacggatcac
      721 ttcgtgtccc tcggtcacat gtgcatcctc tagtagcggt cctgctgagc ctcgacatgt
      781 tgtcgcaaaa ttcgccctgg acccgcccaa cgatttgtcg tcactgtcaa ggtttgacct
      841 gcacttcatt tggggcccac atacaccaaa aaaatgctgc ataattctcg gggcagcaag
      901 tcggttaccc ggccgccgtg ctggaccggg ttgaatggtg cccgtaactt tcggtagagc
      961 ggacggccaa tactcaactt caaggaatct cacccatgcg cgccggcggg gaaccggagt
     1021 tcccttcagt gagcgttatt agttcgccgc tcggtgtgtc gtagatacta gcccctgggg
     1081 cacttttgaa atttgaataa gatttatgta atcagtcttt taggtttgac cggttctgcc
     1141 gcttttttta aaattggatt tgtaataata aaacgcaatt gtttgttatt gtggcgctct
     1201 atcatagatg tcgctataaa cctattcagc acaatatatt gttttcattt taatattgta
     1261 catataagta gtagggtaca atcagtaaat tgaacggaga atattattca taaaaatacg
     1321 atagtaacgg gtgatatatt cattagaatg aaccgaaacc ggcggtaagg atctgagcta
     1381 cacatgctca ggttttttac aacgtgcaca acagaattga aagcaaatat catgcgatca
     1441 taggcttctc gcatatctca ttaaagcagg acaagctcaa gaagtcacca ttgcagagat
     1501 tccgtacacc accaatccag tgatagcagc tccgaacaca acagtcatga tctttccgaa
     1561 ctttgaagat ttctcagggt tcttagcgat ctcgttagcc acaccctcag taagcttagc
     1621 cacctgtgcc tcgtccacgt tcttaagctg tccagatgcc ttcaacttct ggaaagctga
     1681 ttgccacttg ttatttgttg tgtccaagtt tgtaagcttt cctgcctgga tcttagccac
     1741 tgcagcctgc cacttgttct ggtcagccac agggatcttg atgtcgtcga tgttcttgaa
     1801 gttgttagcc ttcaactttt caagagcaga ttgccatttg ttttttccag cagtagtagt
     1861 ctcagccacc ttctcagcgt tctgcaaagc cttctgtgcc ttaagctttt ccaatgcctg
     1921 ttgccactta tttccaccag cagcagccac agcatcatca agctgtcctg ctcccttgat
     1981 cttagccaat gcgtcgtcca cctgtcccac tcccttcatc ttctccaaag cgtcatccaa
     2041 ttgacctgcc ttgaactttg atgccaagtc tgcccatcct gccattgtat cgataattgt
     2101 aaatgtaatt gtaatgttgt ttgttgtttg ttgttgttgg taattgttgt aaaaatgagc
     2161 tcttatactc gagcgtgtcc tctccaaatg aaatgaactt ccttatatag aggaagggtc
     2221 ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca gtggagatgt cacatcaatc
     2281 cacttgcttt gtagacgtgg ttggaacctc ttctttttcc acgatgctcc tcgtgggtgg
     2341 gggtccatct ttgggaccac tgtcggcaga gagatcttga atgatagcct ttcctttatc
     2401 gcaatgatgg catttgtagg agccaccttc cttttctact gtcctttcga tgaagtgaca
     2461 gatagctggg caatggaatc cgaggaggtt tcccgaaatt atcctttgtt gaaaagtctc
     2521 aatagccctt tgatcttctg agactgtatc tttgacattt ttggagtaga ccagagtgtc
     2581 gtgctccacc atgttgacga agattttctt cttgtcattg agtcgtaaaa gactctgtat
     2641 gaactgttcg ccagtcttga cggcgagttc tgttagatcc tcgatttgaa tcttagactc
     2701 catgcatggc cttagattca gtaggaacta cctttttaga gactccaatc tctattactt
     2761 gccttggttt atgaagcaag ccttgaatcg tccatactgg aatagtactt ctgatcttga
     2821 gaaatatgtc tttctctgtg ttcttgatgc aattagtcct gaatcttttg actgcatctt
     2881 taaccttctt gggaaggtat ttgatctcct ggagattgtt actcgggtag atcgtcttga
     2941 tgagacttgc tgcgtaggcc tctctaacca tctgtgggtc agcattcttt ctgaaattga
     3001 agaggctaac cttctcatta tcagtggtga acatagtgtc gtcaccttca ccttcgaact
     3061 tccttcctag atcgtaaaga tagaggaaat cgtccattgt aatctccggg gcaaaggaga
     3121 tctcttttgg ggctggatca ctgctgggcc ttttggttcc tagcgtgagc cagtgggctt
     3181 tttgctttgg tgggcttgtt agggccttag caaagctctt gggcttgagt tgagcttctc
     3241 ctttggggat gaagttcaac ctgtctgttt gctgacttgt tgtgtacgcg tcagctgctg
     3301 ctcttgcctc tgtaatagtg gcaaatttct tgtgtgcaac tccgggaacg ccgtttgttg
     3361 ccgcctttgt acaaccccag tcatcgtata taccggcatg tggaccgtta tacacaacgt
     3421 agtagttgat atgagggtgt tgaatacccg attctgctct gagaggagca actgtgctgt
     3481 taagctcaga tttttgtggg attggaattc ctccaagctt gaattcttgc agcgctccaa
     3541 ttcagcaagc gtcgatctag taacatagat gacaccgcgc gcgataattt atcctagttt
     3601 gcgcgctata ttttgttttc tatcgcgtat taaatgtata attgcgggac tctaatcata
     3661 aaaacccatc tcataaataa cgtcatgcat tacatgttaa ttattacatg cttaacgtaa
     3721 ttcaacagaa attatatgat aatcatcgca agaccggcaa caggattcaa tcttaagaaa
     3781 ctttattgcc aaatgtttga acgatctgct tgactctaga tccagagtcc cgctcaaatc
     3841 tcagtcacag gaagaacagg tctaggagga acaggaagag agaaatcaag ctgccagaat
     3901 cccacatcat gccaatttcc atgtttgaaa ccagcagctc taagcattcc cctaggagca
     3961 taaccaagag cctcatgcat cctcacagaa ggatcattag gaagtccaat cacagcaacc
     4021 acagacttga aaccctgagc ctccaaagac ttcaaaagat gagtgtaaag agtagatcca
     4081 agaccagttc tttgatgcct aggagacacg taaacagtag actcagcagt ccaatcgtaa
     4141 gcattcctag ccttccaagg accagcataa gcaataccag ccacttcacc atccacttca
     4201 gcaacaagcc aaggatacct ctctctaagc ctcacaagat catcagtcca ctcttgtggt
     4261 tcttgaggct cagtcctgaa attcacagta gaagtctcaa tgtaatgatt cacaatagtg
     4321 cacacagcag gcatatcagc ctcagtagcc cttctaatat cagcaggcct tctttcagga
     4381 gacatgcgaa acgatccaga tccggtgcag attatttgga ttgagagtga atatgagact
     4441 ctaattggat accgagggga atttatggaa cgtcagtgga gcattttgac aagaaatatt
     4501 tgctagctga tagtgacctt aggcgacttt tgaacgcgca ataatggttt ctgacgtatg
     4561 tgcttagctc attaaactcc agaaacccgc ggctgagtgg ctccttcaac gttgcggttc
     4621 tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact
     4681 cccttaattc tccgctccgg atccgaattc ggcacagagt gttcaacccc agtacattaa
     4741 aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac cacaatatat
     4801 cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat caccactcga
     4861 tacaggcagc ccatcaacca tgactactgt ccggtccaac agatccgagc aggaaccgta
     4921 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4981 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5041 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5101 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5161 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5221 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5281 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5341 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5401 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5461 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5521 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5581 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5641 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5701 gctaaaacaa ttcatccagt aaaatataat attttatttt ctcccaatca ggcttgatcc
     5761 ccagtaagtc aaaaaatagc tcgacatact gttcttcccc gatatcctcc ctgagcgacc
     5821 ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc gcttctccca agatcaataa
     5881 agccacttac tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg ccgtgggaaa
     5941 agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg cgcggatctt
     6001 taaatggagt gtcctcttcc cagttttcgc aatccacatc ggccagatcg ttattcagta
     6061 agtaatccaa ttcggctaag cggctgtcta agctattcgt atagggacaa tccgatatgt
     6121 cgatggagtg aaagagcctg atgcactccg catacagctc gataatcttt tcagggcttt
     6181 gttcatcttc atactcttcc gagcaaagga cgccatcggc ctcactcatg agcagattgc
     6241 tccagccatc atgccgttca aagtgcagga cctttggaac aggcagcttt ccttccagcc
     6301 atagcatcat gtccttttcc cgttccacat cataggtggt ccctttatac cggctgtccg
     6361 tcatttttaa atataggttt tcattttctc ccaccagctt atatacctta gcaggagaca
     6421 ttccttccgt atcttttacg cagcggtatt tttcgatcag ttttttcaat tccggtgata
     6481 ttctcatttt agccatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6541 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6601 gcactcggag ccttgatgtg ggcgcttttt gcagctgttc ggctgtgcgc tggccagaca
     6661 gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag ttttaggcgg
     6721 aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg ttcccaatgt
     6781 acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt gggttcccaa
     6841 tgtacgtgct atccacagga aagaagcttt ttcgaccttt ttcccctgct agggcaattt
     6901 gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg atcaggttgc
     6961 ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa tcgtactccg
     7021 gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg aactctccgg
     7081 cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc ttgcctgcgg
     7141 cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa aagtaatcgg
     7201 ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat ctcgcggtac atccaatcag
     7261 ctagctcgat ctcgatgtac tccggccgcc cggtttcgct ctttacgatc ttgtagcggc
     7321 taatcaaggc ttcaccctcg gataccgtca ccaggcggcc gttcttggcc ttcttcgtac
     7381 gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt ttctaccagg tcgtctttct
     7441 gctttccgcc atcggctcgc cggcagaact tgagtacgtc cgcaacgtgt ggacggaaca
     7501 cgcggccggg cttgtctccc ttcccttccc ggtatcggtt catggattcg gttagatggg
     7561 aaaccgccat cagtaccagg tcgtaatccc acacactggc catgccggcc ggccctgcgg
     7621 aaacctctac gtgcccgtct ggaagctcgt agcggatcac ctcgccagct cgtcggtcac
     7681 gcttcgacag acggaaaacg gccacgtcca tgatgctgcg actatcgcgg gtgcccacgt
     7741 catagagcat cggaacgaaa aaatctggtt gctcgtcgcc cttgggcggc ttcctaatcg
     7801 acgacgcacc ggctgccggc ggttgccggg attctttgcg gattcgatca gcggccgctt
     7861 gccacgattc accggggcgt gcttctgcct cgatgcgttg ccgctgggcg gcctgcgcgg
     7921 ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc gatttgtacc gggccggatg
     7981 gtttgcgacc gctcacgccg attcctcggg cttgggggtt ccagtgccat tgcagggccg
     8041 gcagacaacc cagccgctta cgcctggcca accgcccgtt cctccacaca tggggcattc
     8101 cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg cctcctttag ccgctaaaat
     8161 tcatctactc atttattcat ttgctcattt actctggtag ctgcgcgatg tattcagata
     8221 gcagctcggt aatggtcttg ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct
     8281 ccgccggcaa ctgaaagttg acccgcttca tggctggcgt gtctgccagg ctggccaacg
     8341 ttgcagcctt gctgctgcgt gcgctcggac ggccggcact tagcgtgttt gtgcttttgc
     8401 tcattttctc tttacctcat taactcaaat gagttttgat ttaatttcag cggccagcgc
     8461 ctggacctcg cgggcagcgt cgccctcggg ttctgattca agaacggttg tgccggcggc
     8521 ggcagtgcct gggtagctca cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta
     8581 cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac
     8641 gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa cc
//
