LOCUS       sequence-342789-        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_20300
FEATURES             Location/Qualifiers
     source          1..8251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    632..656
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(723..1430)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1696..2041)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1733..1753)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3074..3326)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(3926..4108)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(3962..4140)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4303..4327
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4463..5051)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4543..4562)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5222..6016)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6017..6121)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6183..6377
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6443..7516)
                     /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(7945..8251,1..323))
                     /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 cttggctgca ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc
       61 gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg
      121 cggtcaatcg tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc
      181 gcccaatcgc gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg
      241 agttgcaggg cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg
      301 ttaagtacag cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca
      361 tcatatacgc agcgaccgca tgacgcaagc tgttttactc aaatacacat caccttttta
      421 gacggcggcg ctcggtttct tcagcggcca agctggccgg ccaggccgcc agcttggcat
      481 cagacaaacc ggccaggatt tcatgcagcc gcacggctga ctcccgaggg gaaccctgtg
      541 gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc
      601 gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca
      661 ctgatagttt aaacggatca cttcgtgtcc ctcggtcaca tgtgcatcct ctagtagcgg
      721 tcctgctgag cctcgacatg ttgtcgcaaa attcgccctg gacccgccca acgatttgtc
      781 gtcactgtca aggtttgacc tgcacttcat ttggggccca catacaccaa aaaaatgctg
      841 cataattctc ggggcagcaa gtcggttacc cggccgccgt gctggaccgg gttgaatggt
      901 gcccgtaact ttcggtagag cggacggcca atactcaact tcaaggaatc tcacccatgc
      961 gcgccggcgg ggaaccggag ttcccttcag tgagcgttat tagttcgccg ctcggtgtgt
     1021 cgtagatact agcccctggg gcacttttga aatttgaata agatttatgt aatcagtctt
     1081 ttaggtttga ccggttctgc cgcttttttt aaaattggat ttgtaataat aaaacgcaat
     1141 tgtttgttat tgtggcgctc tatcatagat gtcgctataa acctattcag cacaatatat
     1201 tgttttcatt ttaatattgt acatataagt agtagggtac aatcagtaaa ttgaacggag
     1261 aatattattc ataaaaatac gatagtaacg ggtgatatat tcattagaat gaaccgaaac
     1321 cggcggtaag gatctgagct acacatgctc aggtttttta caacgtgcac aacagaattg
     1381 aaagcaaata tcatgcgatc ataggcttct cgcatatctc attaaagcag gacaagctca
     1441 agacttagtc atcttagcct tagccttctt agcagcctcg atcaactttc cgatcttgat
     1501 gtcaggcctc ttcacgatct tgttagtaga ctgagccacg ttcttccaga atcctccgta
     1561 tccgtcaccc cttccagact ggatcacctc ctgagcctcg atcattgtat cgataattgt
     1621 aaatgtaatt gtaatgttgt ttgttgtttg ttgttgttgg taattgttgt aaaaatgagc
     1681 tcttatactc gagcgtgtcc tctccaaatg aaatgaactt ccttatatag aggaagggtc
     1741 ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca gtggagatgt cacatcaatc
     1801 cacttgcttt gtagacgtgg ttggaacctc ttctttttcc acgatgctcc tcgtgggtgg
     1861 gggtccatct ttgggaccac tgtcggcaga gagatcttga atgatagcct ttcctttatc
     1921 gcaatgatgg catttgtagg agccaccttc cttttctact gtcctttcga tgaagtgaca
     1981 gatagctggg caatggaatc cgaggaggtt tcccgaaatt atcctttgtt gaaaagtctc
     2041 aatagccctt tgatcttctg agactgtatc tttgacattt ttggagtaga ccagagtgtc
     2101 gtgctccacc atgttgacga agattttctt cttgtcattg agtcgtaaaa gactctgtat
     2161 gaactgttcg ccagtcttga cggcgagttc tgttagatcc tcgatttgaa tcttagactc
     2221 catgcatggc cttagattca gtaggaacta cctttttaga gactccaatc tctattactt
     2281 gccttggttt atgaagcaag ccttgaatcg tccatactgg aatagtactt ctgatcttga
     2341 gaaatatgtc tttctctgtg ttcttgatgc aattagtcct gaatcttttg actgcatctt
     2401 taaccttctt gggaaggtat ttgatctcct ggagattgtt actcgggtag atcgtcttga
     2461 tgagacttgc tgcgtaggcc tctctaacca tctgtgggtc agcattcttt ctgaaattga
     2521 agaggctaac cttctcatta tcagtggtga acatagtgtc gtcaccttca ccttcgaact
     2581 tccttcctag atcgtaaaga tagaggaaat cgtccattgt aatctccggg gcaaaggaga
     2641 tctcttttgg ggctggatca ctgctgggcc ttttggttcc tagcgtgagc cagtgggctt
     2701 tttgctttgg tgggcttgtt agggccttag caaagctctt gggcttgagt tgagcttctc
     2761 ctttggggat gaagttcaac ctgtctgttt gctgacttgt tgtgtacgcg tcagctgctg
     2821 ctcttgcctc tgtaatagtg gcaaatttct tgtgtgcaac tccgggaacg ccgtttgttg
     2881 ccgcctttgt acaaccccag tcatcgtata taccggcatg tggaccgtta tacacaacgt
     2941 agtagttgat atgagggtgt tgaatacccg attctgctct gagaggagca actgtgctgt
     3001 taagctcaga tttttgtggg attggaattc ctccaagctt gaattcttgc agcgctccaa
     3061 ttcagcaagc gtcgatctag taacatagat gacaccgcgc gcgataattt atcctagttt
     3121 gcgcgctata ttttgttttc tatcgcgtat taaatgtata attgcgggac tctaatcata
     3181 aaaacccatc tcataaataa cgtcatgcat tacatgttaa ttattacatg cttaacgtaa
     3241 ttcaacagaa attatatgat aatcatcgca agaccggcaa caggattcaa tcttaagaaa
     3301 ctttattgcc aaatgtttga acgatctgct tgactctaga tccagagtcc cgctcaaatc
     3361 tcagtcacag gaagaacagg tctaggagga acaggaagag agaaatcaag ctgccagaat
     3421 cccacatcat gccaatttcc atgtttgaaa ccagcagctc taagcattcc cctaggagca
     3481 taaccaagag cctcatgcat cctcacagaa ggatcattag gaagtccaat cacagcaacc
     3541 acagacttga aaccctgagc ctccaaagac ttcaaaagat gagtgtaaag agtagatcca
     3601 agaccagttc tttgatgcct aggagacacg taaacagtag actcagcagt ccaatcgtaa
     3661 gcattcctag ccttccaagg accagcataa gcaataccag ccacttcacc atccacttca
     3721 gcaacaagcc aaggatacct ctctctaagc ctcacaagat catcagtcca ctcttgtggt
     3781 tcttgaggct cagtcctgaa attcacagta gaagtctcaa tgtaatgatt cacaatagtg
     3841 cacacagcag gcatatcagc ctcagtagcc cttctaatat cagcaggcct tctttcagga
     3901 gacatgcgaa acgatccaga tccggtgcag attatttgga ttgagagtga atatgagact
     3961 ctaattggat accgagggga atttatggaa cgtcagtgga gcattttgac aagaaatatt
     4021 tgctagctga tagtgacctt aggcgacttt tgaacgcgca ataatggttt ctgacgtatg
     4081 tgcttagctc attaaactcc agaaacccgc ggctgagtgg ctccttcaac gttgcggttc
     4141 tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact
     4201 cccttaattc tccgctccgg atccgaattc ggcacagagt gttcaacccc agtacattaa
     4261 aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac cacaatatat
     4321 cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat caccactcga
     4381 tacaggcagc ccatcaacca tgactactgt ccggtccaac agatccgagc aggaaccgta
     4441 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4501 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4561 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4621 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4681 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4741 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4801 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4861 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4921 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4981 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5041 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5101 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5161 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5221 gctaaaacaa ttcatccagt aaaatataat attttatttt ctcccaatca ggcttgatcc
     5281 ccagtaagtc aaaaaatagc tcgacatact gttcttcccc gatatcctcc ctgagcgacc
     5341 ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc gcttctccca agatcaataa
     5401 agccacttac tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg ccgtgggaaa
     5461 agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg cgcggatctt
     5521 taaatggagt gtcctcttcc cagttttcgc aatccacatc ggccagatcg ttattcagta
     5581 agtaatccaa ttcggctaag cggctgtcta agctattcgt atagggacaa tccgatatgt
     5641 cgatggagtg aaagagcctg atgcactccg catacagctc gataatcttt tcagggcttt
     5701 gttcatcttc atactcttcc gagcaaagga cgccatcggc ctcactcatg agcagattgc
     5761 tccagccatc atgccgttca aagtgcagga cctttggaac aggcagcttt ccttccagcc
     5821 atagcatcat gtccttttcc cgttccacat cataggtggt ccctttatac cggctgtccg
     5881 tcatttttaa atataggttt tcattttctc ccaccagctt atatacctta gcaggagaca
     5941 ttccttccgt atcttttacg cagcggtatt tttcgatcag ttttttcaat tccggtgata
     6001 ttctcatttt agccatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6061 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6121 gcactcggag ccttgatgtg ggcgcttttt gcagctgttc ggctgtgcgc tggccagaca
     6181 gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag ttttaggcgg
     6241 aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg ttcccaatgt
     6301 acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt gggttcccaa
     6361 tgtacgtgct atccacagga aagaagcttt ttcgaccttt ttcccctgct agggcaattt
     6421 gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg atcaggttgc
     6481 ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa tcgtactccg
     6541 gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg aactctccgg
     6601 cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc ttgcctgcgg
     6661 cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa aagtaatcgg
     6721 ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat ctcgcggtac atccaatcag
     6781 ctagctcgat ctcgatgtac tccggccgcc cggtttcgct ctttacgatc ttgtagcggc
     6841 taatcaaggc ttcaccctcg gataccgtca ccaggcggcc gttcttggcc ttcttcgtac
     6901 gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt ttctaccagg tcgtctttct
     6961 gctttccgcc atcggctcgc cggcagaact tgagtacgtc cgcaacgtgt ggacggaaca
     7021 cgcggccggg cttgtctccc ttcccttccc ggtatcggtt catggattcg gttagatggg
     7081 aaaccgccat cagtaccagg tcgtaatccc acacactggc catgccggcc ggccctgcgg
     7141 aaacctctac gtgcccgtct ggaagctcgt agcggatcac ctcgccagct cgtcggtcac
     7201 gcttcgacag acggaaaacg gccacgtcca tgatgctgcg actatcgcgg gtgcccacgt
     7261 catagagcat cggaacgaaa aaatctggtt gctcgtcgcc cttgggcggc ttcctaatcg
     7321 acgacgcacc ggctgccggc ggttgccggg attctttgcg gattcgatca gcggccgctt
     7381 gccacgattc accggggcgt gcttctgcct cgatgcgttg ccgctgggcg gcctgcgcgg
     7441 ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc gatttgtacc gggccggatg
     7501 gtttgcgacc gctcacgccg attcctcggg cttgggggtt ccagtgccat tgcagggccg
     7561 gcagacaacc cagccgctta cgcctggcca accgcccgtt cctccacaca tggggcattc
     7621 cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg cctcctttag ccgctaaaat
     7681 tcatctactc atttattcat ttgctcattt actctggtag ctgcgcgatg tattcagata
     7741 gcagctcggt aatggtcttg ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct
     7801 ccgccggcaa ctgaaagttg acccgcttca tggctggcgt gtctgccagg ctggccaacg
     7861 ttgcagcctt gctgctgcgt gcgctcggac ggccggcact tagcgtgttt gtgcttttgc
     7921 tcattttctc tttacctcat taactcaaat gagttttgat ttaatttcag cggccagcgc
     7981 ctggacctcg cgggcagcgt cgccctcggg ttctgattca agaacggttg tgccggcggc
     8041 ggcagtgcct gggtagctca cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta
     8101 cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac
     8161 gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa ccagctccac
     8221 caggtcggcg gtggcccata tgtcgtaagg g
//
