LOCUS       sequence-342189-        9166 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 9166)
  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 9166)
  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_04276
FEATURES             Location/Qualifiers
     source          1..9166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(586..1215)
                     /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    1524..1548
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1615..2322)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3503..3848)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3540..3560)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4881..5133)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5733..5915)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5769..5947)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6110..6134
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6270..6858)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6350..6369)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7029..7823)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7824..7928)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7990..8184
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(join(8250..9166,1..157))
                     /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 ggattcgatc agcggccgct tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt
       61 gccgctgggc ggcctgcgcg gccttcaact tctccaccag gtcatcaccc agcgccgcgc
      121 cgatttgtac cgggccggat ggtttgcgac cgctcacgcc gattcctcgg gcttgggggt
      181 tccagtgcca ttgcagggcc ggcagacaac ccagccgctt acgcctggcc aaccgcccgt
      241 tcctccacac atggggcatt ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc
      301 gcctccttta gccgctaaaa ttcatctact catttattca tttgctcatt tactctggta
      361 gctgcgcgat gtattcagat agcagctcgg taatggtctt gccttggcgt accgcgtaca
      421 tcttcagctt ggtgtgatcc tccgccggca actgaaagtt gacccgcttc atggctggcg
      481 tgtctgccag gctggccaac gttgcagcct tgctgctgcg tgcgctcgga cggccggcac
      541 ttagcgtgtt tgtgcttttg ctcattttct ctttacctca ttaactcaaa tgagttttga
      601 tttaatttca gcggccagcg cctggacctc gcgggcagcg tcgccctcgg gttctgattc
      661 aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg tgatacggga
      721 ctcaagaatg ggcagctcgt acccggccag cgcctcggca acctcaccgc cgatgcgcgt
      781 gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg tgacctcaat
      841 gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg
      901 caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt ccgccgcctg
      961 gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat
     1021 cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg ccgcccaatc
     1081 gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg cgagttgcag
     1141 ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct ggttaagtac
     1201 agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg catcatatac
     1261 gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt tagacggcgg
     1321 cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc atcagacaaa
     1381 ccggccagga tttcatgcag ccgcacggct gactcccgag gggaaccctg tggttggcat
     1441 gcacatacaa atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc
     1501 taataaacgc tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt
     1561 ttaaacggat cacttcgtgt ccctcggtca catgtgcatc ctctagtagc ggtcctgctg
     1621 agcctcgaca tgttgtcgca aaattcgccc tggacccgcc caacgatttg tcgtcactgt
     1681 caaggtttga cctgcacttc atttggggcc cacatacacc aaaaaaatgc tgcataattc
     1741 tcggggcagc aagtcggtta cccggccgcc gtgctggacc gggttgaatg gtgcccgtaa
     1801 ctttcggtag agcggacggc caatactcaa cttcaaggaa tctcacccat gcgcgccggc
     1861 ggggaaccgg agttcccttc agtgagcgtt attagttcgc cgctcggtgt gtcgtagata
     1921 ctagcccctg gggcactttt gaaatttgaa taagatttat gtaatcagtc ttttaggttt
     1981 gaccggttct gccgcttttt ttaaaattgg atttgtaata ataaaacgca attgtttgtt
     2041 attgtggcgc tctatcatag atgtcgctat aaacctattc agcacaatat attgttttca
     2101 ttttaatatt gtacatataa gtagtagggt acaatcagta aattgaacgg agaatattat
     2161 tcataaaaat acgatagtaa cgggtgatat attcattaga atgaaccgaa accggcggta
     2221 aggatctgag ctacacatgc tcaggttttt tacaacgtgc acaacagaat tgaaagcaaa
     2281 tatcatgcga tcataggctt ctcgcatatc tcattaaagc aggacaagct catgatcctc
     2341 tagtagcgtg tgagaagtcg tcctggttca ttttcttcct ctgaagtttg ttgaagtatg
     2401 acacccaagt tccgaaagca gggtctgaca aagcgttgtc tccctttcca gacaacctca
     2461 aaagtttgaa agcctcgtga gctggcattc cctccctcaa ccacaaagca gatgccacct
     2521 gagcagcagt agccttaagg ttaccagtaa ttgaagagtg agtaaccatc ttcttaagtg
     2581 tttcatcgtc gtagtatttt gtcataacag atgacataac gttatcagcc tcttctttat
     2641 gcataaagaa ccatgacacc aatcctggag atttcaatgt gtcgatagat tggtcgatat
     2701 caagcctaag catcttgaac acgtcgtctg atgtttttcc cttctccttc caaatcttga
     2761 actgtgtaga ctccaattta acagccattg acaccaagtc agagtctgac ttagccctca
     2821 caagcatctt tggaagttcc tcctcaccgt gcctctttgc caacttcaaa accaagaact
     2881 cgtaaggatc caaagtgtga tgcttagttg cgtaagagat gaaagtttga agtgctggat
     2941 gcttaagaag tcccaactcc ttgtctgcag gcacaagtga gatcttaagc acgtcgtcct
     3001 cagcctttcc tgttgtaagc cacttcttga tctgtgcaaa ctcaagaccc tttgcgatgt
     3061 ccctagttga agcagttgta gatgcagcca tcaacctctt gtgcaatgcg tcctcctcat
     3121 actgctttgc aagtttaagc aacaagaagt cgtatggatc ttccttaagt ttagttttaa
     3181 cgaaagagat ccaagttggc aaaagtgggt tagccaacac gtccacgtcc tccacgttca
     3241 acttcaacaa attatagatg tctcctgcag ttttcttctg agtcacccag ttgtcgaact
     3301 gaatagcctt aactgttgtt gcagcgtgtg aagctccagg agactgagcc tccacaagca
     3361 acttagtaag tgcatcatca ccgttctgag ctgtcaaagc agtcctaaac attgtatcga
     3421 taattgtaaa tgtaattgta atgttgtttg ttgtttgttg ttgttggtaa ttgttgtaaa
     3481 aatgagctct tatactcgag cgtgtcctct ccaaatgaaa tgaacttcct tatatagagg
     3541 aagggtcttg cgaaggatag tgggattgtg cgtcatccct tacgtcagtg gagatgtcac
     3601 atcaatccac ttgctttgta gacgtggttg gaacctcttc tttttccacg atgctcctcg
     3661 tgggtggggg tccatctttg ggaccactgt cggcagagag atcttgaatg atagcctttc
     3721 ctttatcgca atgatggcat ttgtaggagc caccttcctt ttctactgtc ctttcgatga
     3781 agtgacagat agctgggcaa tggaatccga ggaggtttcc cgaaattatc ctttgttgaa
     3841 aagtctcaat agccctttga tcttctgaga ctgtatcttt gacatttttg gagtagacca
     3901 gagtgtcgtg ctccaccatg ttgacgaaga ttttcttctt gtcattgagt cgtaaaagac
     3961 tctgtatgaa ctgttcgcca gtcttgacgg cgagttctgt tagatcctcg atttgaatct
     4021 tagactccat gcatggcctt agattcagta ggaactacct ttttagagac tccaatctct
     4081 attacttgcc ttggtttatg aagcaagcct tgaatcgtcc atactggaat agtacttctg
     4141 atcttgagaa atatgtcttt ctctgtgttc ttgatgcaat tagtcctgaa tcttttgact
     4201 gcatctttaa ccttcttggg aaggtatttg atctcctgga gattgttact cgggtagatc
     4261 gtcttgatga gacttgctgc gtaggcctct ctaaccatct gtgggtcagc attctttctg
     4321 aaattgaaga ggctaacctt ctcattatca gtggtgaaca tagtgtcgtc accttcacct
     4381 tcgaacttcc ttcctagatc gtaaagatag aggaaatcgt ccattgtaat ctccggggca
     4441 aaggagatct cttttggggc tggatcactg ctgggccttt tggttcctag cgtgagccag
     4501 tgggcttttt gctttggtgg gcttgttagg gccttagcaa agctcttggg cttgagttga
     4561 gcttctcctt tggggatgaa gttcaacctg tctgtttgct gacttgttgt gtacgcgtca
     4621 gctgctgctc ttgcctctgt aatagtggca aatttcttgt gtgcaactcc gggaacgccg
     4681 tttgttgccg cctttgtaca accccagtca tcgtatatac cggcatgtgg accgttatac
     4741 acaacgtagt agttgatatg agggtgttga atacccgatt ctgctctgag aggagcaact
     4801 gtgctgttaa gctcagattt ttgtgggatt ggaattcctc caagcttgaa ttcttgcagc
     4861 gctccaattc agcaagcgtc gatctagtaa catagatgac accgcgcgcg ataatttatc
     4921 ctagtttgcg cgctatattt tgttttctat cgcgtattaa atgtataatt gcgggactct
     4981 aatcataaaa acccatctca taaataacgt catgcattac atgttaatta ttacatgctt
     5041 aacgtaattc aacagaaatt atatgataat catcgcaaga ccggcaacag gattcaatct
     5101 taagaaactt tattgccaaa tgtttgaacg atctgcttga ctctagatcc agagtcccgc
     5161 tcaaatctca gtcacaggaa gaacaggtct aggaggaaca ggaagagaga aatcaagctg
     5221 ccagaatccc acatcatgcc aatttccatg tttgaaacca gcagctctaa gcattcccct
     5281 aggagcataa ccaagagcct catgcatcct cacagaagga tcattaggaa gtccaatcac
     5341 agcaaccaca gacttgaaac cctgagcctc caaagacttc aaaagatgag tgtaaagagt
     5401 agatccaaga ccagttcttt gatgcctagg agacacgtaa acagtagact cagcagtcca
     5461 atcgtaagca ttcctagcct tccaaggacc agcataagca ataccagcca cttcaccatc
     5521 cacttcagca acaagccaag gatacctctc tctaagcctc acaagatcat cagtccactc
     5581 ttgtggttct tgaggctcag tcctgaaatt cacagtagaa gtctcaatgt aatgattcac
     5641 aatagtgcac acagcaggca tatcagcctc agtagccctt ctaatatcag caggccttct
     5701 ttcaggagac atgcgaaacg atccagatcc ggtgcagatt atttggattg agagtgaata
     5761 tgagactcta attggatacc gaggggaatt tatggaacgt cagtggagca ttttgacaag
     5821 aaatatttgc tagctgatag tgaccttagg cgacttttga acgcgcaata atggtttctg
     5881 acgtatgtgc ttagctcatt aaactccaga aacccgcggc tgagtggctc cttcaacgtt
     5941 gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa
     6001 cgtgactccc ttaattctcc gctccggatc cgaattcggc acagagtgtt caaccccagt
     6061 acattaaaaa cgtccgcaat gtgttattaa gttgtctaag cgtcaatttg tttacaccac
     6121 aatatatcct gccaccagcc agccaacagc tccccgaccg gcagctcggc acaaaatcac
     6181 cactcgatac aggcagccca tcaaccatga ctactgtccg gtccaacaga tccgagcagg
     6241 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     6301 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     6361 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     6421 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     6481 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     6541 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     6601 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     6661 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     6721 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     6781 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     6841 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6901 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6961 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     7021 tctgacagct aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc
     7081 ttgatcccca gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg
     7141 agcgaccgga cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga
     7201 tcaataaagc cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg
     7261 tgggaaaaga caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc
     7321 ggatctttaa atggagtgtc ctcttcccag ttttcgcaat ccacatcggc cagatcgtta
     7381 ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc tattcgtata gggacaatcc
     7441 gatatgtcga tggagtgaaa gagcctgatg cactccgcat acagctcgat aatcttttca
     7501 gggctttgtt catcttcata ctcttccgag caaaggacgc catcggcctc actcatgagc
     7561 agattgctcc agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct
     7621 tccagccata gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg
     7681 ctgtccgtca tttttaaata taggttttca ttttctccca ccagcttata taccttagca
     7741 ggagacattc cttccgtatc ttttacgcag cggtattttt cgatcagttt tttcaattcc
     7801 ggtgatattc tcattttagc catactcttc ctttttcaat attattgaag catttatcag
     7861 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     7921 gttccgcgca ctcggagcct tgatgtgggc gctttttgca gctgttcggc tgtgcgctgg
     7981 ccagacagtt atgcacaggc caggcgggtt ttaagagttt taataagttt taaagagttt
     8041 taggcggaaa aatcgccttt tttctctttt atatcagtca cttacatgtg tgaccggttc
     8101 ccaatgtacg gctttgggtt cccaatgtac gggttccggt tcccaatgta cggctttggg
     8161 ttcccaatgt acgtgctatc cacaggaaag aagctttttc gacctttttc ccctgctagg
     8221 gcaatttgcc ctagcatctg ctccgtacat taggaaccgg cggatgcttc gccctcgatc
     8281 aggttgcggt agcgcatgac taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg
     8341 tactccggca ggtcatttga cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac
     8401 tctccggcgc tgccactgcg ttcgtagatc gtcttgaaca accatctggc ttctgccttg
     8461 cctgcggcgc ggcgtgccag gcggtagaga aaacggccga tgccgggatc gatcaaaaag
     8521 taatcggggt gaaccgtcag cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc
     8581 caatcagcta gctcgatctc gatgtactcc ggccgcccgg tttcgctctt tacgatcttg
     8641 tagcggctaa tcaaggcttc accctcggat accgtcacca ggcggccgtt cttggccttc
     8701 ttcgtacgct gcatggcaac gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg
     8761 tctttctgct ttccgccatc ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga
     8821 cggaacacgc ggccgggctt gtctcccttc ccttcccggt atcggttcat ggattcggtt
     8881 agatgggaaa ccgccatcag taccaggtcg taatcccaca cactggccat gccggccggc
     8941 cctgcggaaa cctctacgtg cccgtctgga agctcgtagc ggatcacctc gccagctcgt
     9001 cggtcacgct tcgacagacg gaaaacggcc acgtccatga tgctgcgact atcgcgggtg
     9061 cccacgtcat agagcatcgg aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc
     9121 ctaatcgacg acgcaccggc tgccggcggt tgccgggatt ctttgc
//
