LOCUS       sequence-342191-        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_06087
FEATURES             Location/Qualifiers
     source          1..8326
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    704..728
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(795..1502)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1843..2188)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1880..1900)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3221..3473)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4073..4255)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4109..4287)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4450..4474
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4610..5198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4690..4709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5369..6163)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6164..6268)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6330..6524
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6590..7663)
                     /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(8092..8326,1..395))
                     /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 cttccatccg tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat
       61 atgtcgtaag ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac
      121 acagccaagt ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg
      181 gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct
      241 tcccgcacgg ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca
      301 tcggccccgg cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac
      361 ccgcctttct ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat
      421 ttactcatcg catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac
      481 atcacctttt tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg
      541 ccagcttggc atcagacaaa ccggccagga tttcatgcag ccgcacggct gactcccgag
      601 gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct tttcacgccc
      661 ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc cgccaatata
      721 tcctgtcaaa cactgatagt ttaaacggat cacttcgtgt ccctcggtca catgtgcatc
      781 ctctagtagc ggtcctgctg agcctcgaca tgttgtcgca aaattcgccc tggacccgcc
      841 caacgatttg tcgtcactgt caaggtttga cctgcacttc atttggggcc cacatacacc
      901 aaaaaaatgc tgcataattc tcggggcagc aagtcggtta cccggccgcc gtgctggacc
      961 gggttgaatg gtgcccgtaa ctttcggtag agcggacggc caatactcaa cttcaaggaa
     1021 tctcacccat gcgcgccggc ggggaaccgg agttcccttc agtgagcgtt attagttcgc
     1081 cgctcggtgt gtcgtagata ctagcccctg gggcactttt gaaatttgaa taagatttat
     1141 gtaatcagtc ttttaggttt gaccggttct gccgcttttt ttaaaattgg atttgtaata
     1201 ataaaacgca attgtttgtt attgtggcgc tctatcatag atgtcgctat aaacctattc
     1261 agcacaatat attgttttca ttttaatatt gtacatataa gtagtagggt acaatcagta
     1321 aattgaacgg agaatattat tcataaaaat acgatagtaa cgggtgatat attcattaga
     1381 atgaaccgaa accggcggta aggatctgag ctacacatgc tcaggttttt tacaacgtgc
     1441 acaacagaat tgaaagcaaa tatcatgcga tcataggctt ctcgcatatc tcattaaagc
     1501 aggacaagct caaggagtag ccaacctctt cttctcctcc ttcttcttct ccatagcagt
     1561 gtagaaagag tagtatccca agtacaacct ctgcaactgc ttagccacgt tgttcatctc
     1621 tgcctgagac actccaagca ctgttttaac ctgagtagga gtaagcttag ccttgtacca
     1681 agccctgtag ttgatcctcc aagcagccct gtcctcctct ccagcctcag ccaagatagc
     1741 agcgatagac attgtatcga taattgtaaa tgtaattgta atgttgtttg ttgtttgttg
     1801 ttgttggtaa ttgttgtaaa aatgagctct tatactcgag cgtgtcctct ccaaatgaaa
     1861 tgaacttcct tatatagagg aagggtcttg cgaaggatag tgggattgtg cgtcatccct
     1921 tacgtcagtg gagatgtcac atcaatccac ttgctttgta gacgtggttg gaacctcttc
     1981 tttttccacg atgctcctcg tgggtggggg tccatctttg ggaccactgt cggcagagag
     2041 atcttgaatg atagcctttc ctttatcgca atgatggcat ttgtaggagc caccttcctt
     2101 ttctactgtc ctttcgatga agtgacagat agctgggcaa tggaatccga ggaggtttcc
     2161 cgaaattatc ctttgttgaa aagtctcaat agccctttga tcttctgaga ctgtatcttt
     2221 gacatttttg gagtagacca gagtgtcgtg ctccaccatg ttgacgaaga ttttcttctt
     2281 gtcattgagt cgtaaaagac tctgtatgaa ctgttcgcca gtcttgacgg cgagttctgt
     2341 tagatcctcg atttgaatct tagactccat gcatggcctt agattcagta ggaactacct
     2401 ttttagagac tccaatctct attacttgcc ttggtttatg aagcaagcct tgaatcgtcc
     2461 atactggaat agtacttctg atcttgagaa atatgtcttt ctctgtgttc ttgatgcaat
     2521 tagtcctgaa tcttttgact gcatctttaa ccttcttggg aaggtatttg atctcctgga
     2581 gattgttact cgggtagatc gtcttgatga gacttgctgc gtaggcctct ctaaccatct
     2641 gtgggtcagc attctttctg aaattgaaga ggctaacctt ctcattatca gtggtgaaca
     2701 tagtgtcgtc accttcacct tcgaacttcc ttcctagatc gtaaagatag aggaaatcgt
     2761 ccattgtaat ctccggggca aaggagatct cttttggggc tggatcactg ctgggccttt
     2821 tggttcctag cgtgagccag tgggcttttt gctttggtgg gcttgttagg gccttagcaa
     2881 agctcttggg cttgagttga gcttctcctt tggggatgaa gttcaacctg tctgtttgct
     2941 gacttgttgt gtacgcgtca gctgctgctc ttgcctctgt aatagtggca aatttcttgt
     3001 gtgcaactcc gggaacgccg tttgttgccg cctttgtaca accccagtca tcgtatatac
     3061 cggcatgtgg accgttatac acaacgtagt agttgatatg agggtgttga atacccgatt
     3121 ctgctctgag aggagcaact gtgctgttaa gctcagattt ttgtgggatt ggaattcctc
     3181 caagcttgaa ttcttgcagc gctccaattc agcaagcgtc gatctagtaa catagatgac
     3241 accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat cgcgtattaa
     3301 atgtataatt gcgggactct aatcataaaa acccatctca taaataacgt catgcattac
     3361 atgttaatta ttacatgctt aacgtaattc aacagaaatt atatgataat catcgcaaga
     3421 ccggcaacag gattcaatct taagaaactt tattgccaaa tgtttgaacg atctgcttga
     3481 ctctagatcc agagtcccgc tcaaatctca gtcacaggaa gaacaggtct aggaggaaca
     3541 ggaagagaga aatcaagctg ccagaatccc acatcatgcc aatttccatg tttgaaacca
     3601 gcagctctaa gcattcccct aggagcataa ccaagagcct catgcatcct cacagaagga
     3661 tcattaggaa gtccaatcac agcaaccaca gacttgaaac cctgagcctc caaagacttc
     3721 aaaagatgag tgtaaagagt agatccaaga ccagttcttt gatgcctagg agacacgtaa
     3781 acagtagact cagcagtcca atcgtaagca ttcctagcct tccaaggacc agcataagca
     3841 ataccagcca cttcaccatc cacttcagca acaagccaag gatacctctc tctaagcctc
     3901 acaagatcat cagtccactc ttgtggttct tgaggctcag tcctgaaatt cacagtagaa
     3961 gtctcaatgt aatgattcac aatagtgcac acagcaggca tatcagcctc agtagccctt
     4021 ctaatatcag caggccttct ttcaggagac atgcgaaacg atccagatcc ggtgcagatt
     4081 atttggattg agagtgaata tgagactcta attggatacc gaggggaatt tatggaacgt
     4141 cagtggagca ttttgacaag aaatatttgc tagctgatag tgaccttagg cgacttttga
     4201 acgcgcaata atggtttctg acgtatgtgc ttagctcatt aaactccaga aacccgcggc
     4261 tgagtggctc cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc
     4321 gtcatcggcg ggggtcataa cgtgactccc ttaattctcc gctccggatc cgaattcggc
     4381 acagagtgtt caaccccagt acattaaaaa cgtccgcaat gtgttattaa gttgtctaag
     4441 cgtcaatttg tttacaccac aatatatcct gccaccagcc agccaacagc tccccgaccg
     4501 gcagctcggc acaaaatcac cactcgatac aggcagccca tcaaccatga ctactgtccg
     4561 gtccaacaga tccgagcagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4621 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4681 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4741 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4801 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4861 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4921 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4981 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5041 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5101 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5161 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5221 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5281 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5341 gtatatatga gtaaacttgg tctgacagct aaaacaattc atccagtaaa atataatatt
     5401 ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg acatactgtt
     5461 cttccccgat atcctccctg agcgaccgga cgcagaaggc aatgtcatac cacttgtccg
     5521 ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc acaaagatgt
     5581 tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt tccgtcttta
     5641 aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ctcttcccag ttttcgcaat
     5701 ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc
     5761 tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg cactccgcat
     5821 acagctcgat aatcttttca gggctttgtt catcttcata ctcttccgag caaaggacgc
     5881 catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag tgcaggacct
     5941 ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt tccacatcat
     6001 aggtggtccc tttataccgg ctgtccgtca tttttaaata taggttttca ttttctccca
     6061 ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag cggtattttt
     6121 cgatcagttt tttcaattcc ggtgatattc tcattttagc catactcttc ctttttcaat
     6181 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6241 agaaaaataa acaaataggg gttccgcgca ctcggagcct tgatgtgggc gctttttgca
     6301 gctgttcggc tgtgcgctgg ccagacagtt atgcacaggc caggcgggtt ttaagagttt
     6361 taataagttt taaagagttt taggcggaaa aatcgccttt tttctctttt atatcagtca
     6421 cttacatgtg tgaccggttc ccaatgtacg gctttgggtt cccaatgtac gggttccggt
     6481 tcccaatgta cggctttggg ttcccaatgt acgtgctatc cacaggaaag aagctttttc
     6541 gacctttttc ccctgctagg gcaatttgcc ctagcatctg ctccgtacat taggaaccgg
     6601 cggatgcttc gccctcgatc aggttgcggt agcgcatgac taggatcggg ccagcctgcc
     6661 ccgcctcctc cttcaaatcg tactccggca ggtcatttga cccgatcagc ttgcgcacgg
     6721 tgaaacagaa cttcttgaac tctccggcgc tgccactgcg ttcgtagatc gtcttgaaca
     6781 accatctggc ttctgccttg cctgcggcgc ggcgtgccag gcggtagaga aaacggccga
     6841 tgccgggatc gatcaaaaag taatcggggt gaaccgtcag cacgtccggg ttcttgcctt
     6901 ctgtgatctc gcggtacatc caatcagcta gctcgatctc gatgtactcc ggccgcccgg
     6961 tttcgctctt tacgatcttg tagcggctaa tcaaggcttc accctcggat accgtcacca
     7021 ggcggccgtt cttggccttc ttcgtacgct gcatggcaac gtgcgtggtg tttaaccgaa
     7081 tgcaggtttc taccaggtcg tctttctgct ttccgccatc ggctcgccgg cagaacttga
     7141 gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt gtctcccttc ccttcccggt
     7201 atcggttcat ggattcggtt agatgggaaa ccgccatcag taccaggtcg taatcccaca
     7261 cactggccat gccggccggc cctgcggaaa cctctacgtg cccgtctgga agctcgtagc
     7321 ggatcacctc gccagctcgt cggtcacgct tcgacagacg gaaaacggcc acgtccatga
     7381 tgctgcgact atcgcgggtg cccacgtcat agagcatcgg aacgaaaaaa tctggttgct
     7441 cgtcgccctt gggcggcttc ctaatcgacg acgcaccggc tgccggcggt tgccgggatt
     7501 ctttgcggat tcgatcagcg gccgcttgcc acgattcacc ggggcgtgct tctgcctcga
     7561 tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc caccaggtca tcacccagcg
     7621 ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct cacgccgatt cctcgggctt
     7681 gggggttcca gtgccattgc agggccggca gacaacccag ccgcttacgc ctggccaacc
     7741 gcccgttcct ccacacatgg ggcattccac ggcgtcggtg cctggttgtt cttgattttc
     7801 catgccgcct cctttagccg ctaaaattca tctactcatt tattcatttg ctcatttact
     7861 ctggtagctg cgcgatgtat tcagatagca gctcggtaat ggtcttgcct tggcgtaccg
     7921 cgtacatctt cagcttggtg tgatcctccg ccggcaactg aaagttgacc cgcttcatgg
     7981 ctggcgtgtc tgccaggctg gccaacgttg cagccttgct gctgcgtgcg ctcggacggc
     8041 cggcacttag cgtgtttgtg cttttgctca ttttctcttt acctcattaa ctcaaatgag
     8101 ttttgattta atttcagcgg ccagcgcctg gacctcgcgg gcagcgtcgc cctcgggttc
     8161 tgattcaaga acggttgtgc cggcggcggc agtgcctggg tagctcacgc gctgcgtgat
     8221 acgggactca agaatgggca gctcgtaccc ggccagcgcc tcggcaacct caccgccgat
     8281 gcgcgtgcct ttgatcgccc gcgacacgac aaaggccgct tgtagc
//
