LOCUS       sequence-342204-        9334 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 9334)
  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 9334)
  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_09732
FEATURES             Location/Qualifiers
     source          1..9334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(172..801)
                     /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    1110..1134
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1201..1908)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3257..3602)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3294..3314)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4635..4887)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5487..5669)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5523..5701)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5864..5888
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6024..6612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6104..6123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6783..7577)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7578..7682)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7744..7938
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8004..9077)
                     /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 cgtacatctt cagcttggtg tgatcctccg ccggcaactg aaagttgacc cgcttcatgg
       61 ctggcgtgtc tgccaggctg gccaacgttg cagccttgct gctgcgtgcg ctcggacggc
      121 cggcacttag cgtgtttgtg cttttgctca ttttctcttt acctcattaa ctcaaatgag
      181 ttttgattta atttcagcgg ccagcgcctg gacctcgcgg gcagcgtcgc cctcgggttc
      241 tgattcaaga acggttgtgc cggcggcggc agtgcctggg tagctcacgc gctgcgtgat
      301 acgggactca agaatgggca gctcgtaccc ggccagcgcc tcggcaacct caccgccgat
      361 gcgcgtgcct ttgatcgccc gcgacacgac aaaggccgct tgtagccttc catccgtgac
      421 ctcaatgcgc tgcttaacca gctccaccag gtcggcggtg gcccatatgt cgtaagggct
      481 tggctgcacc ggaatcagca cgaagtcggc tgccttgatc gcggacacag ccaagtccgc
      541 cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg ccgatggcct tcacgtcgcg
      601 gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt tgatcttccc gcacggccgc
      661 ccaatcgcgg gcactgccct ggggatcgga atcgactaac agaacatcgg ccccggcgag
      721 ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg cctgacccgc ctttctggtt
      781 aagtacagcg ataaccttca tgcgttcccc ttgcgtattt gtttatttac tcatcgcatc
      841 atatacgcag cgaccgcatg acgcaagctg ttttactcaa atacacatca cctttttaga
      901 cggcggcgct cggtttcttc agcggccaag ctggccggcc aggccgccag cttggcatca
      961 gacaaaccgg ccaggatttc atgcagccgc acggctgact cccgagggga accctgtggt
     1021 tggcatgcac atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccga
     1081 ttattctaat aaacgctctt ttctcttagg tttacccgcc aatatatcct gtcaaacact
     1141 gatagtttaa acggatcact tcgtgtccct cggtcacatg tgcatcctct agtagcggtc
     1201 ctgctgagcc tcgacatgtt gtcgcaaaat tcgccctgga cccgcccaac gatttgtcgt
     1261 cactgtcaag gtttgacctg cacttcattt ggggcccaca tacaccaaaa aaatgctgca
     1321 taattctcgg ggcagcaagt cggttacccg gccgccgtgc tggaccgggt tgaatggtgc
     1381 ccgtaacttt cggtagagcg gacggccaat actcaacttc aaggaatctc acccatgcgc
     1441 gccggcgggg aaccggagtt cccttcagtg agcgttatta gttcgccgct cggtgtgtcg
     1501 tagatactag cccctggggc acttttgaaa tttgaataag atttatgtaa tcagtctttt
     1561 aggtttgacc ggttctgccg ctttttttaa aattggattt gtaataataa aacgcaattg
     1621 tttgttattg tggcgctcta tcatagatgt cgctataaac ctattcagca caatatattg
     1681 ttttcatttt aatattgtac atataagtag tagggtacaa tcagtaaatt gaacggagaa
     1741 tattattcat aaaaatacga tagtaacggg tgatatattc attagaatga accgaaaccg
     1801 gcggtaagga tctgagctac acatgctcag gttttttaca acgtgcacaa cagaattgaa
     1861 agcaaatatc atgcgatcat aggcttctcg catatctcat taaagcagga caagctcagt
     1921 taccaggctt ctgtgattgg taaaaagcag taaatttagc cataatcttc tcctccacag
     1981 ggtttgatgc agtctcagtt ttgtagatca tcttccagaa gttctctgga gtcacaccct
     2041 ctccaatcca agtagcgaac tgctttttct gaagagtagc agcctctgct ccctcaagga
     2101 tcttagcaag catcttgtta cctcccacct gagttgacat agccttaatg aacacctgcc
     2161 tcacctcctt cacttcagcc aactgctcaa gcacgtcgat cttctgatta acaattgaag
     2221 ttggctggtt aatgtcaatg ttaagtgccc taagcataga tccaagtgat tggttgtcct
     2281 tctgcactcc cctctccaca agcaacttct ttgcagccac gtttgtagta gagtcgtcca
     2341 tagctgaaag caacttcctc atcactggtg cgtcaccgaa agcctgagtg tacaacttca
     2401 acactggctt cttctgacct gggttttttg cattgaagtc gtcaacatac ttcatgaacc
     2461 tgttcaagtt agatgctgtg aaagcgtcgt ccacgtcacc tgaaagccta agccactgga
     2521 acacgttagc aggaagctgt tctccccttg tccacctctc gataaggatg ttctcaagtt
     2581 ttgtagcctt attctttgtt gctgctgagt tagtccttgc tccgtaaagc actccaccaa
     2641 ggttgtcctc accacccaca gacttgataa gagtagtaag aagtgtgtcg tcagactgct
     2701 tcacaagctt agcgtactca tcaagcacct ggaacttctg tcccaatccc ctgttatacc
     2761 tgaagttcaa aagcttagaa acagaaccca aagactttcc tgagtccttc catttcatca
     2821 actgctcagt ttgcaacttc tgaatctgct gagccactga tggtgagtca gcagacctct
     2881 ttgctgtcac aagagcagct gccactgcgt cgtcaccgta cttagtagtc aaagttccca
     2941 acaatgacac cttcaccaac ctgtttttgt cgttaatttc cttcaacttt gtaataaggt
     3001 tattgatatt agaagatgca agagtgtcat cgatctcacc gaacctcata gccctagcag
     3061 ccttcatctc tctagccttc aagttcactg ccttgtccac aagtgaagaa gttcttgact
     3121 tcaacttagc agccaagtca gagatagcag ttccaccgat acccattgta tcgataattg
     3181 taaatgtaat tgtaatgttg tttgttgttt gttgttgttg gtaattgttg taaaaatgag
     3241 ctcttatact cgagcgtgtc ctctccaaat gaaatgaact tccttatata gaggaagggt
     3301 cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagatg tcacatcaat
     3361 ccacttgctt tgtagacgtg gttggaacct cttctttttc cacgatgctc ctcgtgggtg
     3421 ggggtccatc tttgggacca ctgtcggcag agagatcttg aatgatagcc tttcctttat
     3481 cgcaatgatg gcatttgtag gagccacctt ccttttctac tgtcctttcg atgaagtgac
     3541 agatagctgg gcaatggaat ccgaggaggt ttcccgaaat tatcctttgt tgaaaagtct
     3601 caatagccct ttgatcttct gagactgtat ctttgacatt tttggagtag accagagtgt
     3661 cgtgctccac catgttgacg aagattttct tcttgtcatt gagtcgtaaa agactctgta
     3721 tgaactgttc gccagtcttg acggcgagtt ctgttagatc ctcgatttga atcttagact
     3781 ccatgcatgg ccttagattc agtaggaact acctttttag agactccaat ctctattact
     3841 tgccttggtt tatgaagcaa gccttgaatc gtccatactg gaatagtact tctgatcttg
     3901 agaaatatgt ctttctctgt gttcttgatg caattagtcc tgaatctttt gactgcatct
     3961 ttaaccttct tgggaaggta tttgatctcc tggagattgt tactcgggta gatcgtcttg
     4021 atgagacttg ctgcgtaggc ctctctaacc atctgtgggt cagcattctt tctgaaattg
     4081 aagaggctaa ccttctcatt atcagtggtg aacatagtgt cgtcaccttc accttcgaac
     4141 ttccttccta gatcgtaaag atagaggaaa tcgtccattg taatctccgg ggcaaaggag
     4201 atctcttttg gggctggatc actgctgggc cttttggttc ctagcgtgag ccagtgggct
     4261 ttttgctttg gtgggcttgt tagggcctta gcaaagctct tgggcttgag ttgagcttct
     4321 cctttgggga tgaagttcaa cctgtctgtt tgctgacttg ttgtgtacgc gtcagctgct
     4381 gctcttgcct ctgtaatagt ggcaaatttc ttgtgtgcaa ctccgggaac gccgtttgtt
     4441 gccgcctttg tacaacccca gtcatcgtat ataccggcat gtggaccgtt atacacaacg
     4501 tagtagttga tatgagggtg ttgaataccc gattctgctc tgagaggagc aactgtgctg
     4561 ttaagctcag atttttgtgg gattggaatt cctccaagct tgaattcttg cagcgctcca
     4621 attcagcaag cgtcgatcta gtaacataga tgacaccgcg cgcgataatt tatcctagtt
     4681 tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat aattgcggga ctctaatcat
     4741 aaaaacccat ctcataaata acgtcatgca ttacatgtta attattacat gcttaacgta
     4801 attcaacaga aattatatga taatcatcgc aagaccggca acaggattca atcttaagaa
     4861 actttattgc caaatgtttg aacgatctgc ttgactctag atccagagtc ccgctcaaat
     4921 ctcagtcaca ggaagaacag gtctaggagg aacaggaaga gagaaatcaa gctgccagaa
     4981 tcccacatca tgccaatttc catgtttgaa accagcagct ctaagcattc ccctaggagc
     5041 ataaccaaga gcctcatgca tcctcacaga aggatcatta ggaagtccaa tcacagcaac
     5101 cacagacttg aaaccctgag cctccaaaga cttcaaaaga tgagtgtaaa gagtagatcc
     5161 aagaccagtt ctttgatgcc taggagacac gtaaacagta gactcagcag tccaatcgta
     5221 agcattccta gccttccaag gaccagcata agcaatacca gccacttcac catccacttc
     5281 agcaacaagc caaggatacc tctctctaag cctcacaaga tcatcagtcc actcttgtgg
     5341 ttcttgaggc tcagtcctga aattcacagt agaagtctca atgtaatgat tcacaatagt
     5401 gcacacagca ggcatatcag cctcagtagc ccttctaata tcagcaggcc ttctttcagg
     5461 agacatgcga aacgatccag atccggtgca gattatttgg attgagagtg aatatgagac
     5521 tctaattgga taccgagggg aatttatgga acgtcagtgg agcattttga caagaaatat
     5581 ttgctagctg atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat
     5641 gtgcttagct cattaaactc cagaaacccg cggctgagtg gctccttcaa cgttgcggtt
     5701 ctgtcagttc caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac
     5761 tcccttaatt ctccgctccg gatccgaatt cggcacagag tgttcaaccc cagtacatta
     5821 aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca ccacaatata
     5881 tcctgccacc agccagccaa cagctccccg accggcagct cggcacaaaa tcaccactcg
     5941 atacaggcag cccatcaacc atgactactg tccggtccaa cagatccgag caggaaccgt
     6001 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     6061 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     6121 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     6181 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     6241 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     6301 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     6361 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     6421 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     6481 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     6541 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     6601 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     6661 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     6721 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     6781 agctaaaaca attcatccag taaaatataa tattttattt tctcccaatc aggcttgatc
     6841 cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc cctgagcgac
     6901 cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc aagatcaata
     6961 aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc gccgtgggaa
     7021 aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc gcgcggatct
     7081 ttaaatggag tgtcctcttc ccagttttcg caatccacat cggccagatc gttattcagt
     7141 aagtaatcca attcggctaa gcggctgtct aagctattcg tatagggaca atccgatatg
     7201 tcgatggagt gaaagagcct gatgcactcc gcatacagct cgataatctt ttcagggctt
     7261 tgttcatctt catactcttc cgagcaaagg acgccatcgg cctcactcat gagcagattg
     7321 ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt tccttccagc
     7381 catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata ccggctgtcc
     7441 gtcattttta aatataggtt ttcattttct cccaccagct tatatacctt agcaggagac
     7501 attccttccg tatcttttac gcagcggtat ttttcgatca gttttttcaa ttccggtgat
     7561 attctcattt tagccatact cttccttttt caatattatt gaagcattta tcagggttat
     7621 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     7681 cgcactcgga gccttgatgt gggcgctttt tgcagctgtt cggctgtgcg ctggccagac
     7741 agttatgcac aggccaggcg ggttttaaga gttttaataa gttttaaaga gttttaggcg
     7801 gaaaaatcgc cttttttctc ttttatatca gtcacttaca tgtgtgaccg gttcccaatg
     7861 tacggctttg ggttcccaat gtacgggttc cggttcccaa tgtacggctt tgggttccca
     7921 atgtacgtgc tatccacagg aaagaagctt tttcgacctt tttcccctgc tagggcaatt
     7981 tgccctagca tctgctccgt acattaggaa ccggcggatg cttcgccctc gatcaggttg
     8041 cggtagcgca tgactaggat cgggccagcc tgccccgcct cctccttcaa atcgtactcc
     8101 ggcaggtcat ttgacccgat cagcttgcgc acggtgaaac agaacttctt gaactctccg
     8161 gcgctgccac tgcgttcgta gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg
     8221 gcgcggcgtg ccaggcggta gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg
     8281 gggtgaaccg tcagcacgtc cgggttcttg ccttctgtga tctcgcggta catccaatca
     8341 gctagctcga tctcgatgta ctccggccgc ccggtttcgc tctttacgat cttgtagcgg
     8401 ctaatcaagg cttcaccctc ggataccgtc accaggcggc cgttcttggc cttcttcgta
     8461 cgctgcatgg caacgtgcgt ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc
     8521 tgctttccgc catcggctcg ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac
     8581 acgcggccgg gcttgtctcc cttcccttcc cggtatcggt tcatggattc ggttagatgg
     8641 gaaaccgcca tcagtaccag gtcgtaatcc cacacactgg ccatgccggc cggccctgcg
     8701 gaaacctcta cgtgcccgtc tggaagctcg tagcggatca cctcgccagc tcgtcggtca
     8761 cgcttcgaca gacggaaaac ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg
     8821 tcatagagca tcggaacgaa aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc
     8881 gacgacgcac cggctgccgg cggttgccgg gattctttgc ggattcgatc agcggccgct
     8941 tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg
     9001 gccttcaact tctccaccag gtcatcaccc agcgccgcgc cgatttgtac cgggccggat
     9061 ggtttgcgac cgctcacgcc gattcctcgg gcttgggggt tccagtgcca ttgcagggcc
     9121 ggcagacaac ccagccgctt acgcctggcc aaccgcccgt tcctccacac atggggcatt
     9181 ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc gcctccttta gccgctaaaa
     9241 ttcatctact catttattca tttgctcatt tactctggta gctgcgcgat gtattcagat
     9301 agcagctcgg taatggtctt gccttggcgt accg
//
