LOCUS       sequence-342190-        8350 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 8350)
  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 8350)
  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_04314
FEATURES             Location/Qualifiers
     source          1..8350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(252..881)
                     /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    1190..1214
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1281..1988)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2353..2698)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2390..2410)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3731..3983)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4583..4765)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4619..4797)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4960..4984
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5120..5708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5200..5219)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5879..6673)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6674..6778)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6840..7034
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7100..8173)
                     /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 tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca gctcggtaat
       61 ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg ccggcaactg
      121 aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg cagccttgct
      181 gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca ttttctcttt
      241 acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg gacctcgcgg
      301 gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc agtgcctggg
      361 tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc ggccagcgcc
      421 tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac aaaggccgct
      481 tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag gtcggcggtg
      541 gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc tgccttgatc
      601 gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg
      661 ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt
      721 tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga atcgactaac
      781 agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg
      841 cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc ttgcgtattt
      901 gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg ttttactcaa
      961 atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag ctggccggcc
     1021 aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc acggctgact
     1081 cccgagggga accctgtggt tggcatgcac atacaaatgg acgaacggat aaaccttttc
     1141 acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg tttacccgcc
     1201 aatatatcct gtcaaacact gatagtttaa acggatcact tcgtgtccct cggtcacatg
     1261 tgcatcctct agtagcggtc ctgctgagcc tcgacatgtt gtcgcaaaat tcgccctgga
     1321 cccgcccaac gatttgtcgt cactgtcaag gtttgacctg cacttcattt ggggcccaca
     1381 tacaccaaaa aaatgctgca taattctcgg ggcagcaagt cggttacccg gccgccgtgc
     1441 tggaccgggt tgaatggtgc ccgtaacttt cggtagagcg gacggccaat actcaacttc
     1501 aaggaatctc acccatgcgc gccggcgggg aaccggagtt cccttcagtg agcgttatta
     1561 gttcgccgct cggtgtgtcg tagatactag cccctggggc acttttgaaa tttgaataag
     1621 atttatgtaa tcagtctttt aggtttgacc ggttctgccg ctttttttaa aattggattt
     1681 gtaataataa aacgcaattg tttgttattg tggcgctcta tcatagatgt cgctataaac
     1741 ctattcagca caatatattg ttttcatttt aatattgtac atataagtag tagggtacaa
     1801 tcagtaaatt gaacggagaa tattattcat aaaaatacga tagtaacggg tgatatattc
     1861 attagaatga accgaaaccg gcggtaagga tctgagctac acatgctcag gttttttaca
     1921 acgtgcacaa cagaattgaa agcaaatatc atgcgatcat aggcttctcg catatctcat
     1981 taaagcagga caagctcaag aattagtctt gtagatccta gctctcactt tgttagagaa
     2041 agtcaccttg tttgggttgt tagcgtgcct cacgatctcg tttcctgcct tcttgtacac
     2101 gttcaagtag ttaagcaaca agtcaggata cttgttctta agcttctccc tgatgtatcc
     2161 cacagtgaag tcgtcccact ctccgaacct cttcaacttg aaagactcag acttcatcat
     2221 cttcttcaac ttagacttag cgaacaatcc caacaagtcc attgtatcga taattgtaaa
     2281 tgtaattgta atgttgtttg ttgtttgttg ttgttggtaa ttgttgtaaa aatgagctct
     2341 tatactcgag cgtgtcctct ccaaatgaaa tgaacttcct tatatagagg aagggtcttg
     2401 cgaaggatag tgggattgtg cgtcatccct tacgtcagtg gagatgtcac atcaatccac
     2461 ttgctttgta gacgtggttg gaacctcttc tttttccacg atgctcctcg tgggtggggg
     2521 tccatctttg ggaccactgt cggcagagag atcttgaatg atagcctttc ctttatcgca
     2581 atgatggcat ttgtaggagc caccttcctt ttctactgtc ctttcgatga agtgacagat
     2641 agctgggcaa tggaatccga ggaggtttcc cgaaattatc ctttgttgaa aagtctcaat
     2701 agccctttga tcttctgaga ctgtatcttt gacatttttg gagtagacca gagtgtcgtg
     2761 ctccaccatg ttgacgaaga ttttcttctt gtcattgagt cgtaaaagac tctgtatgaa
     2821 ctgttcgcca gtcttgacgg cgagttctgt tagatcctcg atttgaatct tagactccat
     2881 gcatggcctt agattcagta ggaactacct ttttagagac tccaatctct attacttgcc
     2941 ttggtttatg aagcaagcct tgaatcgtcc atactggaat agtacttctg atcttgagaa
     3001 atatgtcttt ctctgtgttc ttgatgcaat tagtcctgaa tcttttgact gcatctttaa
     3061 ccttcttggg aaggtatttg atctcctgga gattgttact cgggtagatc gtcttgatga
     3121 gacttgctgc gtaggcctct ctaaccatct gtgggtcagc attctttctg aaattgaaga
     3181 ggctaacctt ctcattatca gtggtgaaca tagtgtcgtc accttcacct tcgaacttcc
     3241 ttcctagatc gtaaagatag aggaaatcgt ccattgtaat ctccggggca aaggagatct
     3301 cttttggggc tggatcactg ctgggccttt tggttcctag cgtgagccag tgggcttttt
     3361 gctttggtgg gcttgttagg gccttagcaa agctcttggg cttgagttga gcttctcctt
     3421 tggggatgaa gttcaacctg tctgtttgct gacttgttgt gtacgcgtca gctgctgctc
     3481 ttgcctctgt aatagtggca aatttcttgt gtgcaactcc gggaacgccg tttgttgccg
     3541 cctttgtaca accccagtca tcgtatatac cggcatgtgg accgttatac acaacgtagt
     3601 agttgatatg agggtgttga atacccgatt ctgctctgag aggagcaact gtgctgttaa
     3661 gctcagattt ttgtgggatt ggaattcctc caagcttgaa ttcttgcagc gctccaattc
     3721 agcaagcgtc gatctagtaa catagatgac accgcgcgcg ataatttatc ctagtttgcg
     3781 cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct aatcataaaa
     3841 acccatctca taaataacgt catgcattac atgttaatta ttacatgctt aacgtaattc
     3901 aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct taagaaactt
     3961 tattgccaaa tgtttgaacg atctgcttga ctctagatcc agagtcccgc tcaaatctca
     4021 gtcacaggaa gaacaggtct aggaggaaca ggaagagaga aatcaagctg ccagaatccc
     4081 acatcatgcc aatttccatg tttgaaacca gcagctctaa gcattcccct aggagcataa
     4141 ccaagagcct catgcatcct cacagaagga tcattaggaa gtccaatcac agcaaccaca
     4201 gacttgaaac cctgagcctc caaagacttc aaaagatgag tgtaaagagt agatccaaga
     4261 ccagttcttt gatgcctagg agacacgtaa acagtagact cagcagtcca atcgtaagca
     4321 ttcctagcct tccaaggacc agcataagca ataccagcca cttcaccatc cacttcagca
     4381 acaagccaag gatacctctc tctaagcctc acaagatcat cagtccactc ttgtggttct
     4441 tgaggctcag tcctgaaatt cacagtagaa gtctcaatgt aatgattcac aatagtgcac
     4501 acagcaggca tatcagcctc agtagccctt ctaatatcag caggccttct ttcaggagac
     4561 atgcgaaacg atccagatcc ggtgcagatt atttggattg agagtgaata tgagactcta
     4621 attggatacc gaggggaatt tatggaacgt cagtggagca ttttgacaag aaatatttgc
     4681 tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc
     4741 ttagctcatt aaactccaga aacccgcggc tgagtggctc cttcaacgtt gcggttctgt
     4801 cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc
     4861 ttaattctcc gctccggatc cgaattcggc acagagtgtt caaccccagt acattaaaaa
     4921 cgtccgcaat gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct
     4981 gccaccagcc agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac
     5041 aggcagccca tcaaccatga ctactgtccg gtccaacaga tccgagcagg aaccgtaaaa
     5101 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     5161 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     5221 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     5281 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     5341 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     5401 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     5461 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     5521 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     5581 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     5641 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     5701 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     5761 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     5821 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct
     5881 aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca
     5941 gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg agcgaccgga
     6001 cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc
     6061 cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga
     6121 caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa
     6181 atggagtgtc ctcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt
     6241 aatccaattc ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga
     6301 tggagtgaaa gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt
     6361 catcttcata ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc
     6421 agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata
     6481 gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca
     6541 tttttaaata taggttttca ttttctccca ccagcttata taccttagca ggagacattc
     6601 cttccgtatc ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc
     6661 tcattttagc catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6721 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6781 ctcggagcct tgatgtgggc gctttttgca gctgttcggc tgtgcgctgg ccagacagtt
     6841 atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa
     6901 aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg
     6961 gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt
     7021 acgtgctatc cacaggaaag aagctttttc gacctttttc ccctgctagg gcaatttgcc
     7081 ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt
     7141 agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca
     7201 ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc
     7261 tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc
     7321 ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt
     7381 gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta
     7441 gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa
     7501 tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct
     7561 gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct
     7621 ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc
     7681 ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa
     7741 ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa
     7801 cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct
     7861 tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat
     7921 agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg
     7981 acgcaccggc tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc
     8041 acgattcacc ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct
     8101 tcaacttctc caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt
     8161 tgcgaccgct cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca
     8221 gacaacccag ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac
     8281 ggcgtcggtg cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca
     8341 tctactcatt
//
