LOCUS       sequence-342751-        8641 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 8641)
  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 8641)
  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_15930
FEATURES             Location/Qualifiers
     source          1..8641
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(306..935)
                     /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    1244..1268
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1335..2042)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2698..3043)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2735..2755)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4076..4328)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4928..5110)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4964..5142)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5305..5329
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5465..6053)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5545..5564)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6224..7018)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7019..7123)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7185..7379
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7445..8518)
                     /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 tgttcttgat tttccatgcc gcctccttta gccgctaaaa ttcatctact catttattca
       61 tttgctcatt tactctggta gctgcgcgat gtattcagat agcagctcgg taatggtctt
      121 gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc tccgccggca actgaaagtt
      181 gacccgcttc atggctggcg tgtctgccag gctggccaac gttgcagcct tgctgctgcg
      241 tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg ctcattttct ctttacctca
      301 ttaactcaaa tgagttttga tttaatttca gcggccagcg cctggacctc gcgggcagcg
      361 tcgccctcgg gttctgattc aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc
      421 acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt acccggccag cgcctcggca
      481 acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc
      541 cttccatccg tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat
      601 atgtcgtaag ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac
      661 acagccaagt ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg
      721 gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct
      781 tcccgcacgg ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca
      841 tcggccccgg cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac
      901 ccgcctttct ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat
      961 ttactcatcg catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac
     1021 atcacctttt tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg
     1081 ccagcttggc atcagacaaa ccggccagga tttcatgcag ccgcacggct gactcccgag
     1141 gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct tttcacgccc
     1201 ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc cgccaatata
     1261 tcctgtcaaa cactgatagt ttaaacggat cacttcgtgt ccctcggtca catgtgcatc
     1321 ctctagtagc ggtcctgctg agcctcgaca tgttgtcgca aaattcgccc tggacccgcc
     1381 caacgatttg tcgtcactgt caaggtttga cctgcacttc atttggggcc cacatacacc
     1441 aaaaaaatgc tgcataattc tcggggcagc aagtcggtta cccggccgcc gtgctggacc
     1501 gggttgaatg gtgcccgtaa ctttcggtag agcggacggc caatactcaa cttcaaggaa
     1561 tctcacccat gcgcgccggc ggggaaccgg agttcccttc agtgagcgtt attagttcgc
     1621 cgctcggtgt gtcgtagata ctagcccctg gggcactttt gaaatttgaa taagatttat
     1681 gtaatcagtc ttttaggttt gaccggttct gccgcttttt ttaaaattgg atttgtaata
     1741 ataaaacgca attgtttgtt attgtggcgc tctatcatag atgtcgctat aaacctattc
     1801 agcacaatat attgttttca ttttaatatt gtacatataa gtagtagggt acaatcagta
     1861 aattgaacgg agaatattat tcataaaaat acgatagtaa cgggtgatat attcattaga
     1921 atgaaccgaa accggcggta aggatctgag ctacacatgc tcaggttttt tacaacgtgc
     1981 acaacagaat tgaaagcaaa tatcatgcga tcataggctt ctcgcatatc tcattaaagc
     2041 aggacaagct cacttctgtg cctcctgcaa cttaaggaac aactggtact ccctagaaag
     2101 ctccttagcc cttgagttgt tctccactcc caacaacttc tccacgtacc agttaggcct
     2161 cttagccttg atccagatcc tgatccttgc ctccatctct gctggagaag gagatggagt
     2221 gatgttagtc aacttgtcag acttcttaac cctctggaag aactggtagt cgaactcagt
     2281 agcatacctc aagtacatcc tgtactcagc gttattcatc aactgcttag cagggatagc
     2341 gttcaatccc aactcagacc aagcaccctg aggtggcacc tcgtcgtaca accacttctt
     2401 cacctgcctt gtctccaagt tgtccacagc cataggcctc ctagagttaa tcttgaactc
     2461 ctcgtagtac ttgtacttag ggttcttctt caactctgta cccttcaatc ccttaagtcc
     2521 aagagcctcc ttcacgtagt cctttgactt tcccatcttt gtccagtagt acacctgagc
     2581 ctgcctgatc aaagatgaga tctccattgt atcgataatt gtaaatgtaa ttgtaatgtt
     2641 gtttgttgtt tgttgttgtt ggtaattgtt gtaaaaatga gctcttatac tcgagcgtgt
     2701 cctctccaaa tgaaatgaac ttccttatat agaggaaggg tcttgcgaag gatagtggga
     2761 ttgtgcgtca tcccttacgt cagtggagat gtcacatcaa tccacttgct ttgtagacgt
     2821 ggttggaacc tcttcttttt ccacgatgct cctcgtgggt gggggtccat ctttgggacc
     2881 actgtcggca gagagatctt gaatgatagc ctttccttta tcgcaatgat ggcatttgta
     2941 ggagccacct tccttttcta ctgtcctttc gatgaagtga cagatagctg ggcaatggaa
     3001 tccgaggagg tttcccgaaa ttatcctttg ttgaaaagtc tcaatagccc tttgatcttc
     3061 tgagactgta tctttgacat ttttggagta gaccagagtg tcgtgctcca ccatgttgac
     3121 gaagattttc ttcttgtcat tgagtcgtaa aagactctgt atgaactgtt cgccagtctt
     3181 gacggcgagt tctgttagat cctcgatttg aatcttagac tccatgcatg gccttagatt
     3241 cagtaggaac taccttttta gagactccaa tctctattac ttgccttggt ttatgaagca
     3301 agccttgaat cgtccatact ggaatagtac ttctgatctt gagaaatatg tctttctctg
     3361 tgttcttgat gcaattagtc ctgaatcttt tgactgcatc tttaaccttc ttgggaaggt
     3421 atttgatctc ctggagattg ttactcgggt agatcgtctt gatgagactt gctgcgtagg
     3481 cctctctaac catctgtggg tcagcattct ttctgaaatt gaagaggcta accttctcat
     3541 tatcagtggt gaacatagtg tcgtcacctt caccttcgaa cttccttcct agatcgtaaa
     3601 gatagaggaa atcgtccatt gtaatctccg gggcaaagga gatctctttt ggggctggat
     3661 cactgctggg ccttttggtt cctagcgtga gccagtgggc tttttgcttt ggtgggcttg
     3721 ttagggcctt agcaaagctc ttgggcttga gttgagcttc tcctttgggg atgaagttca
     3781 acctgtctgt ttgctgactt gttgtgtacg cgtcagctgc tgctcttgcc tctgtaatag
     3841 tggcaaattt cttgtgtgca actccgggaa cgccgtttgt tgccgccttt gtacaacccc
     3901 agtcatcgta tataccggca tgtggaccgt tatacacaac gtagtagttg atatgagggt
     3961 gttgaatacc cgattctgct ctgagaggag caactgtgct gttaagctca gatttttgtg
     4021 ggattggaat tcctccaagc ttgaattctt gcagcgctcc aattcagcaa gcgtcgatct
     4081 agtaacatag atgacaccgc gcgcgataat ttatcctagt ttgcgcgcta tattttgttt
     4141 tctatcgcgt attaaatgta taattgcggg actctaatca taaaaaccca tctcataaat
     4201 aacgtcatgc attacatgtt aattattaca tgcttaacgt aattcaacag aaattatatg
     4261 ataatcatcg caagaccggc aacaggattc aatcttaaga aactttattg ccaaatgttt
     4321 gaacgatctg cttgactcta gatccagagt cccgctcaaa tctcagtcac aggaagaaca
     4381 ggtctaggag gaacaggaag agagaaatca agctgccaga atcccacatc atgccaattt
     4441 ccatgtttga aaccagcagc tctaagcatt cccctaggag cataaccaag agcctcatgc
     4501 atcctcacag aaggatcatt aggaagtcca atcacagcaa ccacagactt gaaaccctga
     4561 gcctccaaag acttcaaaag atgagtgtaa agagtagatc caagaccagt tctttgatgc
     4621 ctaggagaca cgtaaacagt agactcagca gtccaatcgt aagcattcct agccttccaa
     4681 ggaccagcat aagcaatacc agccacttca ccatccactt cagcaacaag ccaaggatac
     4741 ctctctctaa gcctcacaag atcatcagtc cactcttgtg gttcttgagg ctcagtcctg
     4801 aaattcacag tagaagtctc aatgtaatga ttcacaatag tgcacacagc aggcatatca
     4861 gcctcagtag cccttctaat atcagcaggc cttctttcag gagacatgcg aaacgatcca
     4921 gatccggtgc agattatttg gattgagagt gaatatgaga ctctaattgg ataccgaggg
     4981 gaatttatgg aacgtcagtg gagcattttg acaagaaata tttgctagct gatagtgacc
     5041 ttaggcgact tttgaacgcg caataatggt ttctgacgta tgtgcttagc tcattaaact
     5101 ccagaaaccc gcggctgagt ggctccttca acgttgcggt tctgtcagtt ccaaacgtaa
     5161 aacggcttgt cccgcgtcat cggcgggggt cataacgtga ctcccttaat tctccgctcc
     5221 ggatccgaat tcggcacaga gtgttcaacc ccagtacatt aaaaacgtcc gcaatgtgtt
     5281 attaagttgt ctaagcgtca atttgtttac accacaatat atcctgccac cagccagcca
     5341 acagctcccc gaccggcagc tcggcacaaa atcaccactc gatacaggca gcccatcaac
     5401 catgactact gtccggtcca acagatccga gcaggaaccg taaaaaggcc gcgttgctgg
     5461 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5521 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5581 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5641 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     5701 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5761 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5821 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5881 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     5941 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     6001 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     6061 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     6121 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     6181 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagctaaaac aattcatcca
     6241 gtaaaatata atattttatt ttctcccaat caggcttgat ccccagtaag tcaaaaaata
     6301 gctcgacata ctgttcttcc ccgatatcct ccctgagcga ccggacgcag aaggcaatgt
     6361 cataccactt gtccgccctg ccgcttctcc caagatcaat aaagccactt actttgccat
     6421 ctttcacaaa gatgttgctg tctcccaggt cgccgtggga aaagacaagt tcctcttcgg
     6481 gcttttccgt ctttaaaaaa tcatacagct cgcgcggatc tttaaatgga gtgtcctctt
     6541 cccagttttc gcaatccaca tcggccagat cgttattcag taagtaatcc aattcggcta
     6601 agcggctgtc taagctattc gtatagggac aatccgatat gtcgatggag tgaaagagcc
     6661 tgatgcactc cgcatacagc tcgataatct tttcagggct ttgttcatct tcatactctt
     6721 ccgagcaaag gacgccatcg gcctcactca tgagcagatt gctccagcca tcatgccgtt
     6781 caaagtgcag gacctttgga acaggcagct ttccttccag ccatagcatc atgtcctttt
     6841 cccgttccac atcataggtg gtccctttat accggctgtc cgtcattttt aaatataggt
     6901 tttcattttc tcccaccagc ttatatacct tagcaggaga cattccttcc gtatctttta
     6961 cgcagcggta tttttcgatc agttttttca attccggtga tattctcatt ttagccatac
     7021 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7081 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcactcgg agccttgatg
     7141 tgggcgcttt ttgcagctgt tcggctgtgc gctggccaga cagttatgca caggccaggc
     7201 gggttttaag agttttaata agttttaaag agttttaggc ggaaaaatcg ccttttttct
     7261 cttttatatc agtcacttac atgtgtgacc ggttcccaat gtacggcttt gggttcccaa
     7321 tgtacgggtt ccggttccca atgtacggct ttgggttccc aatgtacgtg ctatccacag
     7381 gaaagaagct ttttcgacct ttttcccctg ctagggcaat ttgccctagc atctgctccg
     7441 tacattagga accggcggat gcttcgccct cgatcaggtt gcggtagcgc atgactagga
     7501 tcgggccagc ctgccccgcc tcctccttca aatcgtactc cggcaggtca tttgacccga
     7561 tcagcttgcg cacggtgaaa cagaacttct tgaactctcc ggcgctgcca ctgcgttcgt
     7621 agatcgtctt gaacaaccat ctggcttctg ccttgcctgc ggcgcggcgt gccaggcggt
     7681 agagaaaacg gccgatgccg ggatcgatca aaaagtaatc ggggtgaacc gtcagcacgt
     7741 ccgggttctt gccttctgtg atctcgcggt acatccaatc agctagctcg atctcgatgt
     7801 actccggccg cccggtttcg ctctttacga tcttgtagcg gctaatcaag gcttcaccct
     7861 cggataccgt caccaggcgg ccgttcttgg ccttcttcgt acgctgcatg gcaacgtgcg
     7921 tggtgtttaa ccgaatgcag gtttctacca ggtcgtcttt ctgctttccg ccatcggctc
     7981 gccggcagaa cttgagtacg tccgcaacgt gtggacggaa cacgcggccg ggcttgtctc
     8041 ccttcccttc ccggtatcgg ttcatggatt cggttagatg ggaaaccgcc atcagtacca
     8101 ggtcgtaatc ccacacactg gccatgccgg ccggccctgc ggaaacctct acgtgcccgt
     8161 ctggaagctc gtagcggatc acctcgccag ctcgtcggtc acgcttcgac agacggaaaa
     8221 cggccacgtc catgatgctg cgactatcgc gggtgcccac gtcatagagc atcggaacga
     8281 aaaaatctgg ttgctcgtcg cccttgggcg gcttcctaat cgacgacgca ccggctgccg
     8341 gcggttgccg ggattctttg cggattcgat cagcggccgc ttgccacgat tcaccggggc
     8401 gtgcttctgc ctcgatgcgt tgccgctggg cggcctgcgc ggccttcaac ttctccacca
     8461 ggtcatcacc cagcgccgcg ccgatttgta ccgggccgga tggtttgcga ccgctcacgc
     8521 cgattcctcg ggcttggggg ttccagtgcc attgcagggc cggcagacaa cccagccgct
     8581 tacgcctggc caaccgcccg ttcctccaca catggggcat tccacggcgt cggtgcctgg
     8641 t
//
