LOCUS       sequence-342345-        9406 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 9406)
  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 9406)
  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_15039
FEATURES             Location/Qualifiers
     source          1..9406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(648..1277)
                     /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    1586..1610
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1677..2384)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3805..4150)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3842..3862)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(5183..5435)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(6035..6217)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(6071..6249)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6412..6436
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6572..7160)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6652..6671)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7331..8125)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(8126..8230)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8292..8486
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(8552..9406,1..219))
                     /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 gcccttgggc ggcttcctaa tcgacgacgc accggctgcc ggcggttgcc gggattcttt
       61 gcggattcga tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg
      121 ttgccgctgg gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc
      181 gccgatttgt accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg
      241 gttccagtgc cattgcaggg ccggcagaca acccagccgc ttacgcctgg ccaaccgccc
      301 gttcctccac acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg
      361 ccgcctcctt tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg
      421 tagctgcgcg atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta
      481 catcttcagc ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg
      541 cgtgtctgcc aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc
      601 acttagcgtg tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt
      661 gatttaattt cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat
      721 tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg
      781 gactcaagaa tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc
      841 gtgcctttga tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca
      901 atgcgctgct taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc
      961 tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc
     1021 tggggcgctc cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca
     1081 atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa
     1141 tcgcgggcac tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc
     1201 agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt
     1261 acagcgataa ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat
     1321 acgcagcgac cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc
     1381 ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca
     1441 aaccggccag gatttcatgc agccgcacgg ctgactcccg aggggaaccc tgtggttggc
     1501 atgcacatac aaatggacga acggataaac cttttcacgc ccttttaaat atccgattat
     1561 tctaataaac gctcttttct cttaggttta cccgccaata tatcctgtca aacactgata
     1621 gtttaaacgg atcacttcgt gtccctcggt cacatgtgca tcctctagta gcggtcctgc
     1681 tgagcctcga catgttgtcg caaaattcgc cctggacccg cccaacgatt tgtcgtcact
     1741 gtcaaggttt gacctgcact tcatttgggg cccacataca ccaaaaaaat gctgcataat
     1801 tctcggggca gcaagtcggt tacccggccg ccgtgctgga ccgggttgaa tggtgcccgt
     1861 aactttcggt agagcggacg gccaatactc aacttcaagg aatctcaccc atgcgcgccg
     1921 gcggggaacc ggagttccct tcagtgagcg ttattagttc gccgctcggt gtgtcgtaga
     1981 tactagcccc tggggcactt ttgaaatttg aataagattt atgtaatcag tcttttaggt
     2041 ttgaccggtt ctgccgcttt ttttaaaatt ggatttgtaa taataaaacg caattgtttg
     2101 ttattgtggc gctctatcat agatgtcgct ataaacctat tcagcacaat atattgtttt
     2161 cattttaata ttgtacatat aagtagtagg gtacaatcag taaattgaac ggagaatatt
     2221 attcataaaa atacgatagt aacgggtgat atattcatta gaatgaaccg aaaccggcgg
     2281 taaggatctg agctacacat gctcaggttt tttacaacgt gcacaacaga attgaaagca
     2341 aatatcatgc gatcataggc ttctcgcata tctcattaaa gcaggacaag ctcatcctga
     2401 cttcttttcc ttcaacaatt tagccaatgc atttctgtat ttgttgtaaa gaagcttcct
     2461 cacgtcgtcc ttaccagcac cagacactga aagaaggtag aacaccctca ctgagtctgt
     2521 ctcagttgcc acccacctgt tgatctgtgc gtacaaaagc tgctcagcgt ggtacctagt
     2581 ctttggaacg tgccatgctc tcacgaaagc agtagtaaga gatctctcgt cctcataaat
     2641 ttttgtaaga gaagaaagca agtcacctcc gtcctttggc ttcttacctc tgtagtaatc
     2701 attaaacctt gcccaagaca cgaacaaagg attctcaagc aaaggagtgt tcacatcatc
     2761 caacttaagc aactcgaaaa ccttctttgg agtcaatttt gactcgtacc agttcctgat
     2821 ctgctggatg aacaaatgct gagcgataag agatgaccta ggatttttgt ctgcagcaac
     2881 gatcatctta gccaacacgt cgtcagcata gaacttcttc aacactgaaa taggtgtcat
     2941 ttttgttttg tgatggatca atgaaaagtc gtcagtatac ctaacccatg cagtgaacaa
     3001 tggagactcc aacaatggca ccttagtctc ctgcaatttc aacaaattga aaacgtcgat
     3061 aggttgcttc ttagttttca accaaagagc agtctgctca gcttggatcc taatagcgat
     3121 tttctctgtt gaaggcacgt tcttagctgc gataagcatg tgcacgaaag cctcctcgtc
     3181 tttgtacaaa ggtttcaaaa aagaaaaaag agtagtctct gatgtagggt taaccttatt
     3241 atactcgtca acgaacttag cccaagacac gatctgaggc tgtgtgaaaa gagactcagc
     3301 tgtgtcctcc aacttcaaca acttgaaaac tgagtctggt gtctccttag ccaacaacca
     3361 cctgtagatc tgctcagtct gcaccctttt tgccaaacca gctgtagccc tcacctgctt
     3421 agcagcctcg atgatcttag caagaccgtc atccttgtaa tgagaagtca atgtagcgat
     3481 aactgttgtc ctcttagtag gatttgcttt attgtacaat ttcatataac caatccagtc
     3541 attcaaccta ggatttgcaa gcaagtcgtc tgcagccttg tccaacatca aaagcttgaa
     3601 cacgtcatct gcagactctc ccttcttaag tgcgtcgtcc aactgctgtg acactgatga
     3661 tttaagagag tttgtcaaag tctcgatagc tgatgttcca ggagatggac ccattgtatc
     3721 gataattgta aatgtaattg taatgttgtt tgttgtttgt tgttgttggt aattgttgta
     3781 aaaatgagct cttatactcg agcgtgtcct ctccaaatga aatgaacttc cttatataga
     3841 ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatgtc
     3901 acatcaatcc acttgctttg tagacgtggt tggaacctct tctttttcca cgatgctcct
     3961 cgtgggtggg ggtccatctt tgggaccact gtcggcagag agatcttgaa tgatagcctt
     4021 tcctttatcg caatgatggc atttgtagga gccaccttcc ttttctactg tcctttcgat
     4081 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgaaatta tcctttgttg
     4141 aaaagtctca atagcccttt gatcttctga gactgtatct ttgacatttt tggagtagac
     4201 cagagtgtcg tgctccacca tgttgacgaa gattttcttc ttgtcattga gtcgtaaaag
     4261 actctgtatg aactgttcgc cagtcttgac ggcgagttct gttagatcct cgatttgaat
     4321 cttagactcc atgcatggcc ttagattcag taggaactac ctttttagag actccaatct
     4381 ctattacttg ccttggttta tgaagcaagc cttgaatcgt ccatactgga atagtacttc
     4441 tgatcttgag aaatatgtct ttctctgtgt tcttgatgca attagtcctg aatcttttga
     4501 ctgcatcttt aaccttcttg ggaaggtatt tgatctcctg gagattgtta ctcgggtaga
     4561 tcgtcttgat gagacttgct gcgtaggcct ctctaaccat ctgtgggtca gcattctttc
     4621 tgaaattgaa gaggctaacc ttctcattat cagtggtgaa catagtgtcg tcaccttcac
     4681 cttcgaactt ccttcctaga tcgtaaagat agaggaaatc gtccattgta atctccgggg
     4741 caaaggagat ctcttttggg gctggatcac tgctgggcct tttggttcct agcgtgagcc
     4801 agtgggcttt ttgctttggt gggcttgtta gggccttagc aaagctcttg ggcttgagtt
     4861 gagcttctcc tttggggatg aagttcaacc tgtctgtttg ctgacttgtt gtgtacgcgt
     4921 cagctgctgc tcttgcctct gtaatagtgg caaatttctt gtgtgcaact ccgggaacgc
     4981 cgtttgttgc cgcctttgta caaccccagt catcgtatat accggcatgt ggaccgttat
     5041 acacaacgta gtagttgata tgagggtgtt gaatacccga ttctgctctg agaggagcaa
     5101 ctgtgctgtt aagctcagat ttttgtggga ttggaattcc tccaagcttg aattcttgca
     5161 gcgctccaat tcagcaagcg tcgatctagt aacatagatg acaccgcgcg cgataattta
     5221 tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact
     5281 ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc
     5341 ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat
     5401 cttaagaaac tttattgcca aatgtttgaa cgatctgctt gactctagat ccagagtccc
     5461 gctcaaatct cagtcacagg aagaacaggt ctaggaggaa caggaagaga gaaatcaagc
     5521 tgccagaatc ccacatcatg ccaatttcca tgtttgaaac cagcagctct aagcattccc
     5581 ctaggagcat aaccaagagc ctcatgcatc ctcacagaag gatcattagg aagtccaatc
     5641 acagcaacca cagacttgaa accctgagcc tccaaagact tcaaaagatg agtgtaaaga
     5701 gtagatccaa gaccagttct ttgatgccta ggagacacgt aaacagtaga ctcagcagtc
     5761 caatcgtaag cattcctagc cttccaagga ccagcataag caataccagc cacttcacca
     5821 tccacttcag caacaagcca aggatacctc tctctaagcc tcacaagatc atcagtccac
     5881 tcttgtggtt cttgaggctc agtcctgaaa ttcacagtag aagtctcaat gtaatgattc
     5941 acaatagtgc acacagcagg catatcagcc tcagtagccc ttctaatatc agcaggcctt
     6001 ctttcaggag acatgcgaaa cgatccagat ccggtgcaga ttatttggat tgagagtgaa
     6061 tatgagactc taattggata ccgaggggaa tttatggaac gtcagtggag cattttgaca
     6121 agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc
     6181 tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctgagtggc tccttcaacg
     6241 ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat
     6301 aacgtgactc ccttaattct ccgctccgga tccgaattcg gcacagagtg ttcaacccca
     6361 gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc
     6421 acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc
     6481 accactcgat acaggcagcc catcaaccat gactactgtc cggtccaaca gatccgagca
     6541 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6601 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6661 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6721 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6781 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6841 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6901 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6961 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7021 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7081 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     7141 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7201 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7261 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7321 ggtctgacag ctaaaacaat tcatccagta aaatataata ttttattttc tcccaatcag
     7381 gcttgatccc cagtaagtca aaaaatagct cgacatactg ttcttccccg atatcctccc
     7441 tgagcgaccg gacgcagaag gcaatgtcat accacttgtc cgccctgccg cttctcccaa
     7501 gatcaataaa gccacttact ttgccatctt tcacaaagat gttgctgtct cccaggtcgc
     7561 cgtgggaaaa gacaagttcc tcttcgggct tttccgtctt taaaaaatca tacagctcgc
     7621 gcggatcttt aaatggagtg tcctcttccc agttttcgca atccacatcg gccagatcgt
     7681 tattcagtaa gtaatccaat tcggctaagc ggctgtctaa gctattcgta tagggacaat
     7741 ccgatatgtc gatggagtga aagagcctga tgcactccgc atacagctcg ataatctttt
     7801 cagggctttg ttcatcttca tactcttccg agcaaaggac gccatcggcc tcactcatga
     7861 gcagattgct ccagccatca tgccgttcaa agtgcaggac ctttggaaca ggcagctttc
     7921 cttccagcca tagcatcatg tccttttccc gttccacatc ataggtggtc cctttatacc
     7981 ggctgtccgt catttttaaa tataggtttt cattttctcc caccagctta tataccttag
     8041 caggagacat tccttccgta tcttttacgc agcggtattt ttcgatcagt tttttcaatt
     8101 ccggtgatat tctcatttta gccatactct tcctttttca atattattga agcatttatc
     8161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     8221 gggttccgcg cactcggagc cttgatgtgg gcgctttttg cagctgttcg gctgtgcgct
     8281 ggccagacag ttatgcacag gccaggcggg ttttaagagt tttaataagt tttaaagagt
     8341 tttaggcgga aaaatcgcct tttttctctt ttatatcagt cacttacatg tgtgaccggt
     8401 tcccaatgta cggctttggg ttcccaatgt acgggttccg gttcccaatg tacggctttg
     8461 ggttcccaat gtacgtgcta tccacaggaa agaagctttt tcgacctttt tcccctgcta
     8521 gggcaatttg ccctagcatc tgctccgtac attaggaacc ggcggatgct tcgccctcga
     8581 tcaggttgcg gtagcgcatg actaggatcg ggccagcctg ccccgcctcc tccttcaaat
     8641 cgtactccgg caggtcattt gacccgatca gcttgcgcac ggtgaaacag aacttcttga
     8701 actctccggc gctgccactg cgttcgtaga tcgtcttgaa caaccatctg gcttctgcct
     8761 tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga tcgatcaaaa
     8821 agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc tcgcggtaca
     8881 tccaatcagc tagctcgatc tcgatgtact ccggccgccc ggtttcgctc tttacgatct
     8941 tgtagcggct aatcaaggct tcaccctcgg ataccgtcac caggcggccg ttcttggcct
     9001 tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt tctaccaggt
     9061 cgtctttctg ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc gcaacgtgtg
     9121 gacggaacac gcggccgggc ttgtctccct tcccttcccg gtatcggttc atggattcgg
     9181 ttagatggga aaccgccatc agtaccaggt cgtaatccca cacactggcc atgccggccg
     9241 gccctgcgga aacctctacg tgcccgtctg gaagctcgta gcggatcacc tcgccagctc
     9301 gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat gatgctgcga ctatcgcggg
     9361 tgcccacgtc atagagcatc ggaacgaaaa aatctggttg ctcgtc
//
