LOCUS       sequence-342794-        8326 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 8326)
  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 8326)
  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_22547
FEATURES             Location/Qualifiers
     source          1..8326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    808..832
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(899..1606)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1947..2292)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1984..2004)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3325..3577)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4177..4359)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4213..4391)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4554..4578
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4714..5302)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4794..4813)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5473..6267)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6268..6372)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6434..6628
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6694..7767)
                     /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(8196..8326,1..499))
                     /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 gggactcaag aatgggcagc tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc
       61 gcgtgccttt gatcgcccgc gacacgacaa aggccgcttg tagccttcca tccgtgacct
      121 caatgcgctg cttaaccagc tccaccaggt cggcggtggc ccatatgtcg taagggcttg
      181 gctgcaccgg aatcagcacg aagtcggctg ccttgatcgc ggacacagcc aagtccgccg
      241 cctggggcgc tccgtcgatc actacgaagt cgcgccggcc gatggccttc acgtcgcggt
      301 caatcgtcgg gcggtcgatg ccgacaacgg ttagcggttg atcttcccgc acggccgccc
      361 aatcgcgggc actgccctgg ggatcggaat cgactaacag aacatcggcc ccggcgagtt
      421 gcagggcgcg ggctagatgg gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa
      481 gtacagcgat aaccttcatg cgttcccctt gcgtatttgt ttatttactc atcgcatcat
      541 atacgcagcg accgcatgac gcaagctgtt ttactcaaat acacatcacc tttttagacg
      601 gcggcgctcg gtttcttcag cggccaagct ggccggccag gccgccagct tggcatcaga
      661 caaaccggcc aggatttcat gcagccgcac ggctgactcc cgaggggaac cctgtggttg
      721 gcatgcacat acaaatggac gaacggataa accttttcac gcccttttaa atatccgatt
      781 attctaataa acgctctttt ctcttaggtt tacccgccaa tatatcctgt caaacactga
      841 tagtttaaac ggatcacttc gtgtccctcg gtcacatgtg catcctctag tagcggtcct
      901 gctgagcctc gacatgttgt cgcaaaattc gccctggacc cgcccaacga tttgtcgtca
      961 ctgtcaaggt ttgacctgca cttcatttgg ggcccacata caccaaaaaa atgctgcata
     1021 attctcgggg cagcaagtcg gttacccggc cgccgtgctg gaccgggttg aatggtgccc
     1081 gtaactttcg gtagagcgga cggccaatac tcaacttcaa ggaatctcac ccatgcgcgc
     1141 cggcggggaa ccggagttcc cttcagtgag cgttattagt tcgccgctcg gtgtgtcgta
     1201 gatactagcc cctggggcac ttttgaaatt tgaataagat ttatgtaatc agtcttttag
     1261 gtttgaccgg ttctgccgct ttttttaaaa ttggatttgt aataataaaa cgcaattgtt
     1321 tgttattgtg gcgctctatc atagatgtcg ctataaacct attcagcaca atatattgtt
     1381 ttcattttaa tattgtacat ataagtagta gggtacaatc agtaaattga acggagaata
     1441 ttattcataa aaatacgata gtaacgggtg atatattcat tagaatgaac cgaaaccggc
     1501 ggtaaggatc tgagctacac atgctcaggt tttttacaac gtgcacaaca gaattgaaag
     1561 caaatatcat gcgatcatag gcttctcgca tatctcatta aagcaggaca agctcaccag
     1621 tcgtcagcgt acttagcagc agctccctta agctcagcga acttctcctc agagatacca
     1681 gccttcctca acttcttcaa agcgtcgttc aacctgtcag gtcctccctt cttagcaagc
     1741 ttaggaaggt tagcagtgaa gtcagccctc ttgaacaagt ccaagtagta cttcctagta
     1801 gcctccttag agaacaccct catcaaagag tttcccaagt tctgagtaga agtcattgta
     1861 tcgataattg taaatgtaat tgtaatgttg tttgttgttt gttgttgttg gtaattgttg
     1921 taaaaatgag ctcttatact cgagcgtgtc ctctccaaat gaaatgaact tccttatata
     1981 gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagatg
     2041 tcacatcaat ccacttgctt tgtagacgtg gttggaacct cttctttttc cacgatgctc
     2101 ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag agagatcttg aatgatagcc
     2161 tttcctttat cgcaatgatg gcatttgtag gagccacctt ccttttctac tgtcctttcg
     2221 atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgaaat tatcctttgt
     2281 tgaaaagtct caatagccct ttgatcttct gagactgtat ctttgacatt tttggagtag
     2341 accagagtgt cgtgctccac catgttgacg aagattttct tcttgtcatt gagtcgtaaa
     2401 agactctgta tgaactgttc gccagtcttg acggcgagtt ctgttagatc ctcgatttga
     2461 atcttagact ccatgcatgg ccttagattc agtaggaact acctttttag agactccaat
     2521 ctctattact tgccttggtt tatgaagcaa gccttgaatc gtccatactg gaatagtact
     2581 tctgatcttg agaaatatgt ctttctctgt gttcttgatg caattagtcc tgaatctttt
     2641 gactgcatct ttaaccttct tgggaaggta tttgatctcc tggagattgt tactcgggta
     2701 gatcgtcttg atgagacttg ctgcgtaggc ctctctaacc atctgtgggt cagcattctt
     2761 tctgaaattg aagaggctaa ccttctcatt atcagtggtg aacatagtgt cgtcaccttc
     2821 accttcgaac ttccttccta gatcgtaaag atagaggaaa tcgtccattg taatctccgg
     2881 ggcaaaggag atctcttttg gggctggatc actgctgggc cttttggttc ctagcgtgag
     2941 ccagtgggct ttttgctttg gtgggcttgt tagggcctta gcaaagctct tgggcttgag
     3001 ttgagcttct cctttgggga tgaagttcaa cctgtctgtt tgctgacttg ttgtgtacgc
     3061 gtcagctgct gctcttgcct ctgtaatagt ggcaaatttc ttgtgtgcaa ctccgggaac
     3121 gccgtttgtt gccgcctttg tacaacccca gtcatcgtat ataccggcat gtggaccgtt
     3181 atacacaacg tagtagttga tatgagggtg ttgaataccc gattctgctc tgagaggagc
     3241 aactgtgctg ttaagctcag atttttgtgg gattggaatt cctccaagct tgaattcttg
     3301 cagcgctcca attcagcaag cgtcgatcta gtaacataga tgacaccgcg cgcgataatt
     3361 tatcctagtt tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat aattgcggga
     3421 ctctaatcat aaaaacccat ctcataaata acgtcatgca ttacatgtta attattacat
     3481 gcttaacgta attcaacaga aattatatga taatcatcgc aagaccggca acaggattca
     3541 atcttaagaa actttattgc caaatgtttg aacgatctgc ttgactctag atccagagtc
     3601 ccgctcaaat ctcagtcaca ggaagaacag gtctaggagg aacaggaaga gagaaatcaa
     3661 gctgccagaa tcccacatca tgccaatttc catgtttgaa accagcagct ctaagcattc
     3721 ccctaggagc ataaccaaga gcctcatgca tcctcacaga aggatcatta ggaagtccaa
     3781 tcacagcaac cacagacttg aaaccctgag cctccaaaga cttcaaaaga tgagtgtaaa
     3841 gagtagatcc aagaccagtt ctttgatgcc taggagacac gtaaacagta gactcagcag
     3901 tccaatcgta agcattccta gccttccaag gaccagcata agcaatacca gccacttcac
     3961 catccacttc agcaacaagc caaggatacc tctctctaag cctcacaaga tcatcagtcc
     4021 actcttgtgg ttcttgaggc tcagtcctga aattcacagt agaagtctca atgtaatgat
     4081 tcacaatagt gcacacagca ggcatatcag cctcagtagc ccttctaata tcagcaggcc
     4141 ttctttcagg agacatgcga aacgatccag atccggtgca gattatttgg attgagagtg
     4201 aatatgagac tctaattgga taccgagggg aatttatgga acgtcagtgg agcattttga
     4261 caagaaatat ttgctagctg atagtgacct taggcgactt ttgaacgcgc aataatggtt
     4321 tctgacgtat gtgcttagct cattaaactc cagaaacccg cggctgagtg gctccttcaa
     4381 cgttgcggtt ctgtcagttc caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc
     4441 ataacgtgac tcccttaatt ctccgctccg gatccgaatt cggcacagag tgttcaaccc
     4501 cagtacatta aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca
     4561 ccacaatata tcctgccacc agccagccaa cagctccccg accggcagct cggcacaaaa
     4621 tcaccactcg atacaggcag cccatcaacc atgactactg tccggtccaa cagatccgag
     4681 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4741 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4801 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4861 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4921 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4981 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5041 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5101 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5161 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5221 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5281 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5341 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5401 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5461 ttggtctgac agctaaaaca attcatccag taaaatataa tattttattt tctcccaatc
     5521 aggcttgatc cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc
     5581 cctgagcgac cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc
     5641 aagatcaata aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc
     5701 gccgtgggaa aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc
     5761 gcgcggatct ttaaatggag tgtcctcttc ccagttttcg caatccacat cggccagatc
     5821 gttattcagt aagtaatcca attcggctaa gcggctgtct aagctattcg tatagggaca
     5881 atccgatatg tcgatggagt gaaagagcct gatgcactcc gcatacagct cgataatctt
     5941 ttcagggctt tgttcatctt catactcttc cgagcaaagg acgccatcgg cctcactcat
     6001 gagcagattg ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt
     6061 tccttccagc catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata
     6121 ccggctgtcc gtcattttta aatataggtt ttcattttct cccaccagct tatatacctt
     6181 agcaggagac attccttccg tatcttttac gcagcggtat ttttcgatca gttttttcaa
     6241 ttccggtgat attctcattt tagccatact cttccttttt caatattatt gaagcattta
     6301 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6361 aggggttccg cgcactcgga gccttgatgt gggcgctttt tgcagctgtt cggctgtgcg
     6421 ctggccagac agttatgcac aggccaggcg ggttttaaga gttttaataa gttttaaaga
     6481 gttttaggcg gaaaaatcgc cttttttctc ttttatatca gtcacttaca tgtgtgaccg
     6541 gttcccaatg tacggctttg ggttcccaat gtacgggttc cggttcccaa tgtacggctt
     6601 tgggttccca atgtacgtgc tatccacagg aaagaagctt tttcgacctt tttcccctgc
     6661 tagggcaatt tgccctagca tctgctccgt acattaggaa ccggcggatg cttcgccctc
     6721 gatcaggttg cggtagcgca tgactaggat cgggccagcc tgccccgcct cctccttcaa
     6781 atcgtactcc ggcaggtcat ttgacccgat cagcttgcgc acggtgaaac agaacttctt
     6841 gaactctccg gcgctgccac tgcgttcgta gatcgtcttg aacaaccatc tggcttctgc
     6901 cttgcctgcg gcgcggcgtg ccaggcggta gagaaaacgg ccgatgccgg gatcgatcaa
     6961 aaagtaatcg gggtgaaccg tcagcacgtc cgggttcttg ccttctgtga tctcgcggta
     7021 catccaatca gctagctcga tctcgatgta ctccggccgc ccggtttcgc tctttacgat
     7081 cttgtagcgg ctaatcaagg cttcaccctc ggataccgtc accaggcggc cgttcttggc
     7141 cttcttcgta cgctgcatgg caacgtgcgt ggtgtttaac cgaatgcagg tttctaccag
     7201 gtcgtctttc tgctttccgc catcggctcg ccggcagaac ttgagtacgt ccgcaacgtg
     7261 tggacggaac acgcggccgg gcttgtctcc cttcccttcc cggtatcggt tcatggattc
     7321 ggttagatgg gaaaccgcca tcagtaccag gtcgtaatcc cacacactgg ccatgccggc
     7381 cggccctgcg gaaacctcta cgtgcccgtc tggaagctcg tagcggatca cctcgccagc
     7441 tcgtcggtca cgcttcgaca gacggaaaac ggccacgtcc atgatgctgc gactatcgcg
     7501 ggtgcccacg tcatagagca tcggaacgaa aaaatctggt tgctcgtcgc ccttgggcgg
     7561 cttcctaatc gacgacgcac cggctgccgg cggttgccgg gattctttgc ggattcgatc
     7621 agcggccgct tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt gccgctgggc
     7681 ggcctgcgcg gccttcaact tctccaccag gtcatcaccc agcgccgcgc cgatttgtac
     7741 cgggccggat ggtttgcgac cgctcacgcc gattcctcgg gcttgggggt tccagtgcca
     7801 ttgcagggcc ggcagacaac ccagccgctt acgcctggcc aaccgcccgt tcctccacac
     7861 atggggcatt ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc gcctccttta
     7921 gccgctaaaa ttcatctact catttattca tttgctcatt tactctggta gctgcgcgat
     7981 gtattcagat agcagctcgg taatggtctt gccttggcgt accgcgtaca tcttcagctt
     8041 ggtgtgatcc tccgccggca actgaaagtt gacccgcttc atggctggcg tgtctgccag
     8101 gctggccaac gttgcagcct tgctgctgcg tgcgctcgga cggccggcac ttagcgtgtt
     8161 tgtgcttttg ctcattttct ctttacctca ttaactcaaa tgagttttga tttaatttca
     8221 gcggccagcg cctggacctc gcgggcagcg tcgccctcgg gttctgattc aagaacggtt
     8281 gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg tgatac
//
