LOCUS       sequence-342332-        8689 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 8689)
  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 8689)
  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_13044a
FEATURES             Location/Qualifiers
     source          1..8689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    671..695
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(762..1469)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2173..2518)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2210..2230)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3551..3803)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4403..4585)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4439..4617)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4780..4804
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4940..5528)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5020..5039)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5699..6493)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6494..6598)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6660..6854
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6920..7993)
                     /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(8422..8689,1..362))
                     /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 agctccacca ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac cggaatcagc
       61 acgaagtcgg ctgccttgat cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg
      121 atcactacga agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg
      181 atgccgacaa cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc
      241 tggggatcgg aatcgactaa cagaacatcg gccccggcga gttgcagggc gcgggctaga
      301 tgggttgcga tggtcgtctt gcctgacccg cctttctggt taagtacagc gataaccttc
      361 atgcgttccc cttgcgtatt tgtttattta ctcatcgcat catatacgca gcgaccgcat
      421 gacgcaagct gttttactca aatacacatc acctttttag acggcggcgc tcggtttctt
      481 cagcggccaa gctggccggc caggccgcca gcttggcatc agacaaaccg gccaggattt
      541 catgcagccg cacggctgac tcccgagggg aaccctgtgg ttggcatgca catacaaatg
      601 gacgaacgga taaacctttt cacgcccttt taaatatccg attattctaa taaacgctct
      661 tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta aacggatcac
      721 ttcgtgtccc tcggtcacat gtgcatcctc tagtagcggt cctgctgagc ctcgacatgt
      781 tgtcgcaaaa ttcgccctgg acccgcccaa cgatttgtcg tcactgtcaa ggtttgacct
      841 gcacttcatt tggggcccac atacaccaaa aaaatgctgc ataattctcg gggcagcaag
      901 tcggttaccc ggccgccgtg ctggaccggg ttgaatggtg cccgtaactt tcggtagagc
      961 ggacggccaa tactcaactt caaggaatct cacccatgcg cgccggcggg gaaccggagt
     1021 tcccttcagt gagcgttatt agttcgccgc tcggtgtgtc gtagatacta gcccctgggg
     1081 cacttttgaa atttgaataa gatttatgta atcagtcttt taggtttgac cggttctgcc
     1141 gcttttttta aaattggatt tgtaataata aaacgcaatt gtttgttatt gtggcgctct
     1201 atcatagatg tcgctataaa cctattcagc acaatatatt gttttcattt taatattgta
     1261 catataagta gtagggtaca atcagtaaat tgaacggaga atattattca taaaaatacg
     1321 atagtaacgg gtgatatatt cattagaatg aaccgaaacc ggcggtaagg atctgagcta
     1381 cacatgctca ggttttttac aacgtgcaca acagaattga aagcaaatat catgcgatca
     1441 taggcttctc gcatatctca ttaaagcagg acaagctcaa gtcaccattg cgttgattcc
     1501 gtacaccacc aatccagtga tagcagctcc gaacacgaaa gtcatgatct ttccgaactt
     1561 tgaagatttc tcagggttct tagcgatctc gttagccaca ccctcagtaa gcttagccac
     1621 ctgtgcctcg tccacgttct taagctgtcc agatgccttc aacttctgga aagctgattg
     1681 ccacttgtta tttgttgtgt ccaagtttgt aagctttcct gcctggatct tagccactgc
     1741 agcctgccac ttgttctggt cagccacagg gatcttgatg tcgtcgatgt tcttgaagtt
     1801 gttagccttc aacttttcaa gagcagattg ccatttgttt tttccagcag tagtagtctc
     1861 agccaccttc tcagcgttct gcaaagcctt ctgtgcctta agcttttcca atgcctgttg
     1921 ccacttattt ccaccagcag cagccacagc atcatcaagc tgtcctgctc ccttgatctt
     1981 agccaatgcg tcgtccacct gtcccactcc cttcatcttc tccaaagcgt catccaattg
     2041 acctgccttg aactttgatg ccaagtctgc ccatcctgcc attgtatcga taattgtaaa
     2101 tgtaattgta atgttgtttg ttgtttgttg ttgttggtaa ttgttgtaaa aatgagctct
     2161 tatactcgag cgtgtcctct ccaaatgaaa tgaacttcct tatatagagg aagggtcttg
     2221 cgaaggatag tgggattgtg cgtcatccct tacgtcagtg gagatgtcac atcaatccac
     2281 ttgctttgta gacgtggttg gaacctcttc tttttccacg atgctcctcg tgggtggggg
     2341 tccatctttg ggaccactgt cggcagagag atcttgaatg atagcctttc ctttatcgca
     2401 atgatggcat ttgtaggagc caccttcctt ttctactgtc ctttcgatga agtgacagat
     2461 agctgggcaa tggaatccga ggaggtttcc cgaaattatc ctttgttgaa aagtctcaat
     2521 agccctttga tcttctgaga ctgtatcttt gacatttttg gagtagacca gagtgtcgtg
     2581 ctccaccatg ttgacgaaga ttttcttctt gtcattgagt cgtaaaagac tctgtatgaa
     2641 ctgttcgcca gtcttgacgg cgagttctgt tagatcctcg atttgaatct tagactccat
     2701 gcatggcctt agattcagta ggaactacct ttttagagac tccaatctct attacttgcc
     2761 ttggtttatg aagcaagcct tgaatcgtcc atactggaat agtacttctg atcttgagaa
     2821 atatgtcttt ctctgtgttc ttgatgcaat tagtcctgaa tcttttgact gcatctttaa
     2881 ccttcttggg aaggtatttg atctcctgga gattgttact cgggtagatc gtcttgatga
     2941 gacttgctgc gtaggcctct ctaaccatct gtgggtcagc attctttctg aaattgaaga
     3001 ggctaacctt ctcattatca gtggtgaaca tagtgtcgtc accttcacct tcgaacttcc
     3061 ttcctagatc gtaaagatag aggaaatcgt ccattgtaat ctccggggca aaggagatct
     3121 cttttggggc tggatcactg ctgggccttt tggttcctag cgtgagccag tgggcttttt
     3181 gctttggtgg gcttgttagg gccttagcaa agctcttggg cttgagttga gcttctcctt
     3241 tggggatgaa gttcaacctg tctgtttgct gacttgttgt gtacgcgtca gctgctgctc
     3301 ttgcctctgt aatagtggca aatttcttgt gtgcaactcc gggaacgccg tttgttgccg
     3361 cctttgtaca accccagtca tcgtatatac cggcatgtgg accgttatac acaacgtagt
     3421 agttgatatg agggtgttga atacccgatt ctgctctgag aggagcaact gtgctgttaa
     3481 gctcagattt ttgtgggatt ggaattcctc caagcttgaa ttcttgcagc gctccaattc
     3541 agcaagcgtc gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg
     3601 cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa
     3661 acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc
     3721 aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt
     3781 tattgccaaa tgtttgaacg atctgcttga ctctagatcc agagtcccgc tcaaatctca
     3841 gtcacaggaa gaacaggtct aggaggaaca ggaagagaga aatcaagctg ccagaatccc
     3901 acatcatgcc aatttccatg tttgaaacca gcagctctaa gcattcccct aggagcataa
     3961 ccaagagcct catgcatcct cacagaagga tcattaggaa gtccaatcac agcaaccaca
     4021 gacttgaaac cctgagcctc caaagacttc aaaagatgag tgtaaagagt agatccaaga
     4081 ccagttcttt gatgcctagg agacacgtaa acagtagact cagcagtcca atcgtaagca
     4141 ttcctagcct tccaaggacc agcataagca ataccagcca cttcaccatc cacttcagca
     4201 acaagccaag gatacctctc tctaagcctc acaagatcat cagtccactc ttgtggttct
     4261 tgaggctcag tcctgaaatt cacagtagaa gtctcaatgt aatgattcac aatagtgcac
     4321 acagcaggca tatcagcctc agtagccctt ctaatatcag caggccttct ttcaggagac
     4381 atgcgaaacg atccagatcc ggtgcagatt atttggattg agagtgaata tgagactcta
     4441 attggatacc gaggggaatt tatggaacgt cagtggagca ttttgacaag aaatatttgc
     4501 tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc
     4561 ttagctcatt aaactccaga aacccgcggc tgagtggctc cttcaacgtt gcggttctgt
     4621 cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc
     4681 ttaattctcc gctccggatc cgaattcggc acagagtgtt caaccccagt acattaaaaa
     4741 cgtccgcaat gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct
     4801 gccaccagcc agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac
     4861 aggcagccca tcaaccatga ctactgtccg gtccaacaga tccgagcagg aaccgtaaaa
     4921 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     4981 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5041 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5101 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5161 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5221 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5281 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5341 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5401 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5461 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5521 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5581 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5641 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct
     5701 aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca
     5761 gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg agcgaccgga
     5821 cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc
     5881 cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga
     5941 caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa
     6001 atggagtgtc ctcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt
     6061 aatccaattc ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga
     6121 tggagtgaaa gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt
     6181 catcttcata ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc
     6241 agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata
     6301 gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca
     6361 tttttaaata taggttttca ttttctccca ccagcttata taccttagca ggagacattc
     6421 cttccgtatc ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc
     6481 tcattttagc catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6541 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6601 ctcggagcct tgatgtgggc gctttttgca gctgttcggc tgtgcgctgg ccagacagtt
     6661 atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa
     6721 aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg
     6781 gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt
     6841 acgtgctatc cacaggaaag aagctttttc gacctttttc ccctgctagg gcaatttgcc
     6901 ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt
     6961 agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca
     7021 ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc
     7081 tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc
     7141 ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt
     7201 gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta
     7261 gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa
     7321 tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct
     7381 gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct
     7441 ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc
     7501 ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa
     7561 ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa
     7621 cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct
     7681 tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat
     7741 agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg
     7801 acgcaccggc tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc
     7861 acgattcacc ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct
     7921 tcaacttctc caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt
     7981 tgcgaccgct cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca
     8041 gacaacccag ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac
     8101 ggcgtcggtg cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca
     8161 tctactcatt tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca
     8221 gctcggtaat ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg
     8281 ccggcaactg aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg
     8341 cagccttgct gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca
     8401 ttttctcttt acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg
     8461 gacctcgcgg gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc
     8521 agtgcctggg tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc
     8581 ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac
     8641 aaaggccgct tgtagccttc catccgtgac ctcaatgcgc tgcttaacc
//
