LOCUS       sequence-342341-        8353 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 8353)
  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 8353)
  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_14371
FEATURES             Location/Qualifiers
     source          1..8353
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1323..1952)
                     /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    2261..2285
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(2352..3059)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3427..3772)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3464..3484)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4805..5057)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5657..5839)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5693..5871)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6034..6058
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6194..6782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6274..6293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6953..7747)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7748..7852)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7914..8108
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(8174..8353,1..894))
                     /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 cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg cggcgtgcca ggcggtagag
       61 aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg tgaaccgtca gcacgtccgg
      121 gttcttgcct tctgtgatct cgcggtacat ccaatcagct agctcgatct cgatgtactc
      181 cggccgcccg gtttcgctct ttacgatctt gtagcggcta atcaaggctt caccctcgga
      241 taccgtcacc aggcggccgt tcttggcctt cttcgtacgc tgcatggcaa cgtgcgtggt
      301 gtttaaccga atgcaggttt ctaccaggtc gtctttctgc tttccgccat cggctcgccg
      361 gcagaacttg agtacgtccg caacgtgtgg acggaacacg cggccgggct tgtctccctt
      421 cccttcccgg tatcggttca tggattcggt tagatgggaa accgccatca gtaccaggtc
      481 gtaatcccac acactggcca tgccggccgg ccctgcggaa acctctacgt gcccgtctgg
      541 aagctcgtag cggatcacct cgccagctcg tcggtcacgc ttcgacagac ggaaaacggc
      601 cacgtccatg atgctgcgac tatcgcgggt gcccacgtca tagagcatcg gaacgaaaaa
      661 atctggttgc tcgtcgccct tgggcggctt cctaatcgac gacgcaccgg ctgccggcgg
      721 ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc cacgattcac cggggcgtgc
      781 ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc ttcaacttct ccaccaggtc
      841 atcacccagc gccgcgccga tttgtaccgg gccggatggt ttgcgaccgc tcacgccgat
      901 tcctcgggct tgggggttcc agtgccattg cagggccggc agacaaccca gccgcttacg
      961 cctggccaac cgcccgttcc tccacacatg gggcattcca cggcgtcggt gcctggttgt
     1021 tcttgatttt ccatgccgcc tcctttagcc gctaaaattc atctactcat ttattcattt
     1081 gctcatttac tctggtagct gcgcgatgta ttcagatagc agctcggtaa tggtcttgcc
     1141 ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc gccggcaact gaaagttgac
     1201 ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt gcagccttgc tgctgcgtgc
     1261 gctcggacgg ccggcactta gcgtgtttgt gcttttgctc attttctctt tacctcatta
     1321 actcaaatga gttttgattt aatttcagcg gccagcgcct ggacctcgcg ggcagcgtcg
     1381 ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg gtagctcacg
     1441 cgctgcgtga tacgggactc aagaatgggc agctcgtacc cggccagcgc ctcggcaacc
     1501 tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc ttgtagcctt
     1561 ccatccgtga cctcaatgcg ctgcttaacc agctccacca ggtcggcggt ggcccatatg
     1621 tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg ctgccttgat cgcggacaca
     1681 gccaagtccg ccgcctgggg cgctccgtcg atcactacga agtcgcgccg gccgatggcc
     1741 ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg ttgatcttcc
     1801 cgcacggccg cccaatcgcg ggcactgccc tggggatcgg aatcgactaa cagaacatcg
     1861 gccccggcga gttgcagggc gcgggctaga tgggttgcga tggtcgtctt gcctgacccg
     1921 cctttctggt taagtacagc gataaccttc atgcgttccc cttgcgtatt tgtttattta
     1981 ctcatcgcat catatacgca gcgaccgcat gacgcaagct gttttactca aatacacatc
     2041 acctttttag acggcggcgc tcggtttctt cagcggccaa gctggccggc caggccgcca
     2101 gcttggcatc agacaaaccg gccaggattt catgcagccg cacggctgac tcccgagggg
     2161 aaccctgtgg ttggcatgca catacaaatg gacgaacgga taaacctttt cacgcccttt
     2221 taaatatccg attattctaa taaacgctct tttctcttag gtttacccgc caatatatcc
     2281 tgtcaaacac tgatagttta aacggatcac ttcgtgtccc tcggtcacat gtgcatcctc
     2341 tagtagcggt cctgctgagc ctcgacatgt tgtcgcaaaa ttcgccctgg acccgcccaa
     2401 cgatttgtcg tcactgtcaa ggtttgacct gcacttcatt tggggcccac atacaccaaa
     2461 aaaatgctgc ataattctcg gggcagcaag tcggttaccc ggccgccgtg ctggaccggg
     2521 ttgaatggtg cccgtaactt tcggtagagc ggacggccaa tactcaactt caaggaatct
     2581 cacccatgcg cgccggcggg gaaccggagt tcccttcagt gagcgttatt agttcgccgc
     2641 tcggtgtgtc gtagatacta gcccctgggg cacttttgaa atttgaataa gatttatgta
     2701 atcagtcttt taggtttgac cggttctgcc gcttttttta aaattggatt tgtaataata
     2761 aaacgcaatt gtttgttatt gtggcgctct atcatagatg tcgctataaa cctattcagc
     2821 acaatatatt gttttcattt taatattgta catataagta gtagggtaca atcagtaaat
     2881 tgaacggaga atattattca taaaaatacg atagtaacgg gtgatatatt cattagaatg
     2941 aaccgaaacc ggcggtaagg atctgagcta cacatgctca ggttttttac aacgtgcaca
     3001 acagaattga aagcaaatat catgcgatca taggcttctc gcatatctca ttaaagcagg
     3061 acaagctcag tatccagtca atcccaagtg catcatgtaa gagttgtaga tctggttgta
     3121 cttagctccc tgagtctttc tagaagccaa cttcaagaat gtgtcgatct ggtccaaagt
     3181 caccctgttg tgctgccacc acatgtaagc agcgtcagcc aacttgtcgt ttcccatcaa
     3241 ctgctttgca agcttctttg cgtcagccct ctcagtcaaa gcagccacgt tgaacatctc
     3301 ctcgttcaag ttagccaagt tgaagttagg agccattgta tcgataattg taaatgtaat
     3361 tgtaatgttg tttgttgttt gttgttgttg gtaattgttg taaaaatgag ctcttatact
     3421 cgagcgtgtc ctctccaaat gaaatgaact tccttatata gaggaagggt cttgcgaagg
     3481 atagtgggat tgtgcgtcat cccttacgtc agtggagatg tcacatcaat ccacttgctt
     3541 tgtagacgtg gttggaacct cttctttttc cacgatgctc ctcgtgggtg ggggtccatc
     3601 tttgggacca ctgtcggcag agagatcttg aatgatagcc tttcctttat cgcaatgatg
     3661 gcatttgtag gagccacctt ccttttctac tgtcctttcg atgaagtgac agatagctgg
     3721 gcaatggaat ccgaggaggt ttcccgaaat tatcctttgt tgaaaagtct caatagccct
     3781 ttgatcttct gagactgtat ctttgacatt tttggagtag accagagtgt cgtgctccac
     3841 catgttgacg aagattttct tcttgtcatt gagtcgtaaa agactctgta tgaactgttc
     3901 gccagtcttg acggcgagtt ctgttagatc ctcgatttga atcttagact ccatgcatgg
     3961 ccttagattc agtaggaact acctttttag agactccaat ctctattact tgccttggtt
     4021 tatgaagcaa gccttgaatc gtccatactg gaatagtact tctgatcttg agaaatatgt
     4081 ctttctctgt gttcttgatg caattagtcc tgaatctttt gactgcatct ttaaccttct
     4141 tgggaaggta tttgatctcc tggagattgt tactcgggta gatcgtcttg atgagacttg
     4201 ctgcgtaggc ctctctaacc atctgtgggt cagcattctt tctgaaattg aagaggctaa
     4261 ccttctcatt atcagtggtg aacatagtgt cgtcaccttc accttcgaac ttccttccta
     4321 gatcgtaaag atagaggaaa tcgtccattg taatctccgg ggcaaaggag atctcttttg
     4381 gggctggatc actgctgggc cttttggttc ctagcgtgag ccagtgggct ttttgctttg
     4441 gtgggcttgt tagggcctta gcaaagctct tgggcttgag ttgagcttct cctttgggga
     4501 tgaagttcaa cctgtctgtt tgctgacttg ttgtgtacgc gtcagctgct gctcttgcct
     4561 ctgtaatagt ggcaaatttc ttgtgtgcaa ctccgggaac gccgtttgtt gccgcctttg
     4621 tacaacccca gtcatcgtat ataccggcat gtggaccgtt atacacaacg tagtagttga
     4681 tatgagggtg ttgaataccc gattctgctc tgagaggagc aactgtgctg ttaagctcag
     4741 atttttgtgg gattggaatt cctccaagct tgaattcttg cagcgctcca attcagcaag
     4801 cgtcgatcta gtaacataga tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat
     4861 attttgtttt ctatcgcgta ttaaatgtat aattgcggga ctctaatcat aaaaacccat
     4921 ctcataaata acgtcatgca ttacatgtta attattacat gcttaacgta attcaacaga
     4981 aattatatga taatcatcgc aagaccggca acaggattca atcttaagaa actttattgc
     5041 caaatgtttg aacgatctgc ttgactctag atccagagtc ccgctcaaat ctcagtcaca
     5101 ggaagaacag gtctaggagg aacaggaaga gagaaatcaa gctgccagaa tcccacatca
     5161 tgccaatttc catgtttgaa accagcagct ctaagcattc ccctaggagc ataaccaaga
     5221 gcctcatgca tcctcacaga aggatcatta ggaagtccaa tcacagcaac cacagacttg
     5281 aaaccctgag cctccaaaga cttcaaaaga tgagtgtaaa gagtagatcc aagaccagtt
     5341 ctttgatgcc taggagacac gtaaacagta gactcagcag tccaatcgta agcattccta
     5401 gccttccaag gaccagcata agcaatacca gccacttcac catccacttc agcaacaagc
     5461 caaggatacc tctctctaag cctcacaaga tcatcagtcc actcttgtgg ttcttgaggc
     5521 tcagtcctga aattcacagt agaagtctca atgtaatgat tcacaatagt gcacacagca
     5581 ggcatatcag cctcagtagc ccttctaata tcagcaggcc ttctttcagg agacatgcga
     5641 aacgatccag atccggtgca gattatttgg attgagagtg aatatgagac tctaattgga
     5701 taccgagggg aatttatgga acgtcagtgg agcattttga caagaaatat ttgctagctg
     5761 atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat gtgcttagct
     5821 cattaaactc cagaaacccg cggctgagtg gctccttcaa cgttgcggtt ctgtcagttc
     5881 caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt
     5941 ctccgctccg gatccgaatt cggcacagag tgttcaaccc cagtacatta aaaacgtccg
     6001 caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc
     6061 agccagccaa cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag
     6121 cccatcaacc atgactactg tccggtccaa cagatccgag caggaaccgt aaaaaggccg
     6181 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6241 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6301 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6361 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6421 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6481 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6541 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6601 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     6661 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6721 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6781 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6841 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6901 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agctaaaaca
     6961 attcatccag taaaatataa tattttattt tctcccaatc aggcttgatc cccagtaagt
     7021 caaaaaatag ctcgacatac tgttcttccc cgatatcctc cctgagcgac cggacgcaga
     7081 aggcaatgtc ataccacttg tccgccctgc cgcttctccc aagatcaata aagccactta
     7141 ctttgccatc tttcacaaag atgttgctgt ctcccaggtc gccgtgggaa aagacaagtt
     7201 cctcttcggg cttttccgtc tttaaaaaat catacagctc gcgcggatct ttaaatggag
     7261 tgtcctcttc ccagttttcg caatccacat cggccagatc gttattcagt aagtaatcca
     7321 attcggctaa gcggctgtct aagctattcg tatagggaca atccgatatg tcgatggagt
     7381 gaaagagcct gatgcactcc gcatacagct cgataatctt ttcagggctt tgttcatctt
     7441 catactcttc cgagcaaagg acgccatcgg cctcactcat gagcagattg ctccagccat
     7501 catgccgttc aaagtgcagg acctttggaa caggcagctt tccttccagc catagcatca
     7561 tgtccttttc ccgttccaca tcataggtgg tccctttata ccggctgtcc gtcattttta
     7621 aatataggtt ttcattttct cccaccagct tatatacctt agcaggagac attccttccg
     7681 tatcttttac gcagcggtat ttttcgatca gttttttcaa ttccggtgat attctcattt
     7741 tagccatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7801 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcactcgga
     7861 gccttgatgt gggcgctttt tgcagctgtt cggctgtgcg ctggccagac agttatgcac
     7921 aggccaggcg ggttttaaga gttttaataa gttttaaaga gttttaggcg gaaaaatcgc
     7981 cttttttctc ttttatatca gtcacttaca tgtgtgaccg gttcccaatg tacggctttg
     8041 ggttcccaat gtacgggttc cggttcccaa tgtacggctt tgggttccca atgtacgtgc
     8101 tatccacagg aaagaagctt tttcgacctt tttcccctgc tagggcaatt tgccctagca
     8161 tctgctccgt acattaggaa ccggcggatg cttcgccctc gatcaggttg cggtagcgca
     8221 tgactaggat cgggccagcc tgccccgcct cctccttcaa atcgtactcc ggcaggtcat
     8281 ttgacccgat cagcttgcgc acggtgaaac agaacttctt gaactctccg gcgctgccac
     8341 tgcgttcgta gat
//
