LOCUS       sequence-342329-        9916 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 9916)
  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 9916)
  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_12057
FEATURES             Location/Qualifiers
     source          1..9916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(3..181)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    344..368
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(504..1092)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(584..603)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1263..2057)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(2058..2162)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2224..2418
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2484..3557)
                     /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(3986..4615)
                     /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    4924..4948
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(5015..5722)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(7653..7998)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(7690..7710)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(9031..9283)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(join(9883..9916,1..149))
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
ORIGIN
        1 tctaattgga taccgagggg aatttatgga acgtcagtgg agcattttga caagaaatat
       61 ttgctagctg atagtgacct taggcgactt ttgaacgcgc aataatggtt tctgacgtat
      121 gtgcttagct cattaaactc cagaaacccg cggctgagtg gctccttcaa cgttgcggtt
      181 ctgtcagttc caaacgtaaa acggcttgtc ccgcgtcatc ggcgggggtc ataacgtgac
      241 tcccttaatt ctccgctccg gatccgaatt cggcacagag tgttcaaccc cagtacatta
      301 aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca ccacaatata
      361 tcctgccacc agccagccaa cagctccccg accggcagct cggcacaaaa tcaccactcg
      421 atacaggcag cccatcaacc atgactactg tccggtccaa cagatccgag caggaaccgt
      481 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      541 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      601 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      661 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      721 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      781 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      841 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      901 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
      961 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1021 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1081 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1141 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1201 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     1261 agctaaaaca attcatccag taaaatataa tattttattt tctcccaatc aggcttgatc
     1321 cccagtaagt caaaaaatag ctcgacatac tgttcttccc cgatatcctc cctgagcgac
     1381 cggacgcaga aggcaatgtc ataccacttg tccgccctgc cgcttctccc aagatcaata
     1441 aagccactta ctttgccatc tttcacaaag atgttgctgt ctcccaggtc gccgtgggaa
     1501 aagacaagtt cctcttcggg cttttccgtc tttaaaaaat catacagctc gcgcggatct
     1561 ttaaatggag tgtcctcttc ccagttttcg caatccacat cggccagatc gttattcagt
     1621 aagtaatcca attcggctaa gcggctgtct aagctattcg tatagggaca atccgatatg
     1681 tcgatggagt gaaagagcct gatgcactcc gcatacagct cgataatctt ttcagggctt
     1741 tgttcatctt catactcttc cgagcaaagg acgccatcgg cctcactcat gagcagattg
     1801 ctccagccat catgccgttc aaagtgcagg acctttggaa caggcagctt tccttccagc
     1861 catagcatca tgtccttttc ccgttccaca tcataggtgg tccctttata ccggctgtcc
     1921 gtcattttta aatataggtt ttcattttct cccaccagct tatatacctt agcaggagac
     1981 attccttccg tatcttttac gcagcggtat ttttcgatca gttttttcaa ttccggtgat
     2041 attctcattt tagccatact cttccttttt caatattatt gaagcattta tcagggttat
     2101 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     2161 cgcactcgga gccttgatgt gggcgctttt tgcagctgtt cggctgtgcg ctggccagac
     2221 agttatgcac aggccaggcg ggttttaaga gttttaataa gttttaaaga gttttaggcg
     2281 gaaaaatcgc cttttttctc ttttatatca gtcacttaca tgtgtgaccg gttcccaatg
     2341 tacggctttg ggttcccaat gtacgggttc cggttcccaa tgtacggctt tgggttccca
     2401 atgtacgtgc tatccacagg aaagaagctt tttcgacctt tttcccctgc tagggcaatt
     2461 tgccctagca tctgctccgt acattaggaa ccggcggatg cttcgccctc gatcaggttg
     2521 cggtagcgca tgactaggat cgggccagcc tgccccgcct cctccttcaa atcgtactcc
     2581 ggcaggtcat ttgacccgat cagcttgcgc acggtgaaac agaacttctt gaactctccg
     2641 gcgctgccac tgcgttcgta gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg
     2701 gcgcggcgtg ccaggcggta gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg
     2761 gggtgaaccg tcagcacgtc cgggttcttg ccttctgtga tctcgcggta catccaatca
     2821 gctagctcga tctcgatgta ctccggccgc ccggtttcgc tctttacgat cttgtagcgg
     2881 ctaatcaagg cttcaccctc ggataccgtc accaggcggc cgttcttggc cttcttcgta
     2941 cgctgcatgg caacgtgcgt ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc
     3001 tgctttccgc catcggctcg ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac
     3061 acgcggccgg gcttgtctcc cttcccttcc cggtatcggt tcatggattc ggttagatgg
     3121 gaaaccgcca tcagtaccag gtcgtaatcc cacacactgg ccatgccggc cggccctgcg
     3181 gaaacctcta cgtgcccgtc tggaagctcg tagcggatca cctcgccagc tcgtcggtca
     3241 cgcttcgaca gacggaaaac ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg
     3301 tcatagagca tcggaacgaa aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc
     3361 gacgacgcac cggctgccgg cggttgccgg gattctttgc ggattcgatc agcggccgct
     3421 tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg
     3481 gccttcaact tctccaccag gtcatcaccc agcgccgcgc cgatttgtac cgggccggat
     3541 ggtttgcgac cgctcacgcc gattcctcgg gcttgggggt tccagtgcca ttgcagggcc
     3601 ggcagacaac ccagccgctt acgcctggcc aaccgcccgt tcctccacac atggggcatt
     3661 ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc gcctccttta gccgctaaaa
     3721 ttcatctact catttattca tttgctcatt tactctggta gctgcgcgat gtattcagat
     3781 agcagctcgg taatggtctt gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc
     3841 tccgccggca actgaaagtt gacccgcttc atggctggcg tgtctgccag gctggccaac
     3901 gttgcagcct tgctgctgcg tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg
     3961 ctcattttct ctttacctca ttaactcaaa tgagttttga tttaatttca gcggccagcg
     4021 cctggacctc gcgggcagcg tcgccctcgg gttctgattc aagaacggtt gtgccggcgg
     4081 cggcagtgcc tgggtagctc acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt
     4141 acccggccag cgcctcggca acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca
     4201 cgacaaaggc cgcttgtagc cttccatccg tgacctcaat gcgctgctta accagctcca
     4261 ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg caccggaatc agcacgaagt
     4321 cggctgcctt gatcgcggac acagccaagt ccgccgcctg gggcgctccg tcgatcacta
     4381 cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga
     4441 caacggttag cggttgatct tcccgcacgg ccgcccaatc gcgggcactg ccctggggat
     4501 cggaatcgac taacagaaca tcggccccgg cgagttgcag ggcgcgggct agatgggttg
     4561 cgatggtcgt cttgcctgac ccgcctttct ggttaagtac agcgataacc ttcatgcgtt
     4621 ccccttgcgt atttgtttat ttactcatcg catcatatac gcagcgaccg catgacgcaa
     4681 gctgttttac tcaaatacac atcacctttt tagacggcgg cgctcggttt cttcagcggc
     4741 caagctggcc ggccaggccg ccagcttggc atcagacaaa ccggccagga tttcatgcag
     4801 ccgcacggct gactcccgag gggaaccctg tggttggcat gcacatacaa atggacgaac
     4861 ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc tcttttctct
     4921 taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaacggat cacttcgtgt
     4981 ccctcggtca catgtgcatc ctctagtagc ggtcctgctg agcctcgaca tgttgtcgca
     5041 aaattcgccc tggacccgcc caacgatttg tcgtcactgt caaggtttga cctgcacttc
     5101 atttggggcc cacatacacc aaaaaaatgc tgcataattc tcggggcagc aagtcggtta
     5161 cccggccgcc gtgctggacc gggttgaatg gtgcccgtaa ctttcggtag agcggacggc
     5221 caatactcaa cttcaaggaa tctcacccat gcgcgccggc ggggaaccgg agttcccttc
     5281 agtgagcgtt attagttcgc cgctcggtgt gtcgtagata ctagcccctg gggcactttt
     5341 gaaatttgaa taagatttat gtaatcagtc ttttaggttt gaccggttct gccgcttttt
     5401 ttaaaattgg atttgtaata ataaaacgca attgtttgtt attgtggcgc tctatcatag
     5461 atgtcgctat aaacctattc agcacaatat attgttttca ttttaatatt gtacatataa
     5521 gtagtagggt acaatcagta aattgaacgg agaatattat tcataaaaat acgatagtaa
     5581 cgggtgatat attcattaga atgaaccgaa accggcggta aggatctgag ctacacatgc
     5641 tcaggttttt tacaacgtgc acaacagaat tgaaagcaaa tatcatgcga tcataggctt
     5701 ctcgcatatc tcattaaagc aggacaagct cagttcttct tgtacaaaga caaagcgtgc
     5761 tccacgtcag atgccaactt ctcagtagtt gggttagact tagcagtctt aatcatctgg
     5821 aacaactcat cctcatcgta gtacttcaag aaagttctga tcatgtcagg cttctgcttg
     5881 tagtatgtag ccatatactg gatccagatt tccaagaaag gtgatgcaaa aagtgtcttt
     5941 cctgcaagct ccaagtccaa accctcgaag atctggtctg gagtccttgt ctttctagac
     6001 acgttccagt tcttaagcaa ctccctctcc attctctttg caatcttctc tgtagaagaa
     6061 accttcatac cagagatgat cattttcacc aatgacttat cgtcgtggtg acccctcaca
     6121 gttttcatca tagtcctctg caagtctggg aaagtcaagt aaaaatcgtc aaggtactgg
     6181 atccaagtag cgaactgtgg tgatgccaac aagtcgtcac ccttctctgc caacctcaac
     6241 attgagaaag tgtcagatgg caactttccc accatcctct ccaccaactc gatgttcaca
     6301 ccagttgcag ccaacatagt ccagatagta gtctcctttc catggttctc tctgttataa
     6361 gcagacaagt attttcccca agtgatcacc aatgggtttg tgaaaagctc gtccttctcc
     6421 tgagcaaggt ggaaaacgtt tgtgaaaagg tattcaggag tccatccctt ctgcatccac
     6481 ctctggaaaa gctctgactg aagcttctta gctgcacctg cagtgtcctt cacatgtgtt
     6541 gccttatcca acaatgaaac aaggttaagc tctccgtagt ttgctgccat tgtaccaata
     6601 agagttgcct tgtactttgg gtttgcagcg ttatactcct tcatgtaatt aatccaagtc
     6661 ttaagctgag ggtttgaaag cacgtcgtca tcgaacttgt ccaacttcaa ctgcttgaag
     6721 aatgcgtcag gagttccctt agccaaccac ttctgctcag tctcaagttc cctaataagc
     6781 tttgcagatc ctggcacgtt cttagcagac tcaagcatct tgataagtcc gtcagagtcg
     6841 tagtgtgctg ccaaaacaga catcaaagat gttcttttat cagggttagc agtattcaaa
     6901 tcatccacgt aagaaaccca cttcataaac tgagggtggt cgaaaagctt ctcacctgct
     6961 ttagacaaag acaagaactg aaacacatcc ctaggtgact tctctgtgta aagccatctt
     7021 ttgatctgct cgtcctgcaa gtttgctgca agcttagctg ttgaaggaga tctcattgcg
     7081 tcgaaaagca tagcagtaag cttgtcatct ccgaagtaat ttgtcataac agcaatccta
     7141 gagattctag tatttttaga gttgaaatcg tcaaggtatg taagccaagt tgcgaactga
     7201 gggttctcca aaagcttgtc tcctgccttg tccaatgcca aaaacttgaa cacgtccctt
     7261 ggcctctttc cagcgtcaag ccacaattga atttgttcat tgtgcaattt accagccaca
     7321 ttagctgtag aaggtgtttt catagccctg atcaagattg ttgaaagtgc ctcgtcaccg
     7381 tagtgtgtag tcaagatagg gatagactca tccacctttg acattgtttt ctctttaaag
     7441 tttgttgcgt aagaaagcca agctgttgca agtgggttct ccaacaattt atcacctgct
     7501 tttgaaagtt ccaaaagctt aaacacgtca gctggagctt tcttgttgtc caaccagtac
     7561 attgtatcga taattgtaaa tgtaattgta atgttgtttg ttgtttgttg ttgttggtaa
     7621 ttgttgtaaa aatgagctct tatactcgag cgtgtcctct ccaaatgaaa tgaacttcct
     7681 tatatagagg aagggtcttg cgaaggatag tgggattgtg cgtcatccct tacgtcagtg
     7741 gagatgtcac atcaatccac ttgctttgta gacgtggttg gaacctcttc tttttccacg
     7801 atgctcctcg tgggtggggg tccatctttg ggaccactgt cggcagagag atcttgaatg
     7861 atagcctttc ctttatcgca atgatggcat ttgtaggagc caccttcctt ttctactgtc
     7921 ctttcgatga agtgacagat agctgggcaa tggaatccga ggaggtttcc cgaaattatc
     7981 ctttgttgaa aagtctcaat agccctttga tcttctgaga ctgtatcttt gacatttttg
     8041 gagtagacca gagtgtcgtg ctccaccatg ttgacgaaga ttttcttctt gtcattgagt
     8101 cgtaaaagac tctgtatgaa ctgttcgcca gtcttgacgg cgagttctgt tagatcctcg
     8161 atttgaatct tagactccat gcatggcctt agattcagta ggaactacct ttttagagac
     8221 tccaatctct attacttgcc ttggtttatg aagcaagcct tgaatcgtcc atactggaat
     8281 agtacttctg atcttgagaa atatgtcttt ctctgtgttc ttgatgcaat tagtcctgaa
     8341 tcttttgact gcatctttaa ccttcttggg aaggtatttg atctcctgga gattgttact
     8401 cgggtagatc gtcttgatga gacttgctgc gtaggcctct ctaaccatct gtgggtcagc
     8461 attctttctg aaattgaaga ggctaacctt ctcattatca gtggtgaaca tagtgtcgtc
     8521 accttcacct tcgaacttcc ttcctagatc gtaaagatag aggaaatcgt ccattgtaat
     8581 ctccggggca aaggagatct cttttggggc tggatcactg ctgggccttt tggttcctag
     8641 cgtgagccag tgggcttttt gctttggtgg gcttgttagg gccttagcaa agctcttggg
     8701 cttgagttga gcttctcctt tggggatgaa gttcaacctg tctgtttgct gacttgttgt
     8761 gtacgcgtca gctgctgctc ttgcctctgt aatagtggca aatttcttgt gtgcaactcc
     8821 gggaacgccg tttgttgccg cctttgtaca accccagtca tcgtatatac cggcatgtgg
     8881 accgttatac acaacgtagt agttgatatg agggtgttga atacccgatt ctgctctgag
     8941 aggagcaact gtgctgttaa gctcagattt ttgtgggatt ggaattcctc caagcttgaa
     9001 ttcttgcagc gctccaattc agcaagcgtc gatctagtaa catagatgac accgcgcgcg
     9061 ataatttatc ctagtttgcg cgctatattt tgttttctat cgcgtattaa atgtataatt
     9121 gcgggactct aatcataaaa acccatctca taaataacgt catgcattac atgttaatta
     9181 ttacatgctt aacgtaattc aacagaaatt atatgataat catcgcaaga ccggcaacag
     9241 gattcaatct taagaaactt tattgccaaa tgtttgaacg atctgcttga ctctagatcc
     9301 agagtcccgc tcaaatctca gtcacaggaa gaacaggtct aggaggaaca ggaagagaga
     9361 aatcaagctg ccagaatccc acatcatgcc aatttccatg tttgaaacca gcagctctaa
     9421 gcattcccct aggagcataa ccaagagcct catgcatcct cacagaagga tcattaggaa
     9481 gtccaatcac agcaaccaca gacttgaaac cctgagcctc caaagacttc aaaagatgag
     9541 tgtaaagagt agatccaaga ccagttcttt gatgcctagg agacacgtaa acagtagact
     9601 cagcagtcca atcgtaagca ttcctagcct tccaaggacc agcataagca ataccagcca
     9661 cttcaccatc cacttcagca acaagccaag gatacctctc tctaagcctc acaagatcat
     9721 cagtccactc ttgtggttct tgaggctcag tcctgaaatt cacagtagaa gtctcaatgt
     9781 aatgattcac aatagtgcac acagcaggca tatcagcctc agtagccctt ctaatatcag
     9841 caggccttct ttcaggagac atgcgaaacg atccagatcc ggtgcagatt atttggattg
     9901 agagtgaata tgagac
//
