LOCUS       sequence-342547-        9469 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 9469)
  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 9469)
  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_15278
FEATURES             Location/Qualifiers
     source          1..9469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(330..959)
                     /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    1268..1292
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1359..2066)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3550..3895)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3587..3607)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4928..5180)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5780..5962)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5816..5994)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6157..6181
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6317..6905)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6397..6416)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7076..7870)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7871..7975)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8037..8231
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8297..9370)
                     /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 cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct
       61 aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc
      121 agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg
      181 atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc
      241 caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct
      301 tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc
      361 agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg
      421 gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc
      481 tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc
      541 gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc
      601 tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg
      661 aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc
      721 actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg
      781 ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg
      841 ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg
      901 gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg
      961 cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg accgcatgac
     1021 gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag
     1081 cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat
     1141 gcagccgcac ggctgactcc cgaggggaac cctgtggttg gcatgcacat acaaatggac
     1201 gaacggataa accttttcac gcccttttaa atatccgatt attctaataa acgctctttt
     1261 ctcttaggtt tacccgccaa tatatcctgt caaacactga tagtttaaac ggatcacttc
     1321 gtgtccctcg gtcacatgtg catcctctag tagcggtcct gctgagcctc gacatgttgt
     1381 cgcaaaattc gccctggacc cgcccaacga tttgtcgtca ctgtcaaggt ttgacctgca
     1441 cttcatttgg ggcccacata caccaaaaaa atgctgcata attctcgggg cagcaagtcg
     1501 gttacccggc cgccgtgctg gaccgggttg aatggtgccc gtaactttcg gtagagcgga
     1561 cggccaatac tcaacttcaa ggaatctcac ccatgcgcgc cggcggggaa ccggagttcc
     1621 cttcagtgag cgttattagt tcgccgctcg gtgtgtcgta gatactagcc cctggggcac
     1681 ttttgaaatt tgaataagat ttatgtaatc agtcttttag gtttgaccgg ttctgccgct
     1741 ttttttaaaa ttggatttgt aataataaaa cgcaattgtt tgttattgtg gcgctctatc
     1801 atagatgtcg ctataaacct attcagcaca atatattgtt ttcattttaa tattgtacat
     1861 ataagtagta gggtacaatc agtaaattga acggagaata ttattcataa aaatacgata
     1921 gtaacgggtg atatattcat tagaatgaac cgaaaccggc ggtaaggatc tgagctacac
     1981 atgctcaggt tttttacaac gtgcacaaca gaattgaaag caaatatcat gcgatcatag
     2041 gcttctcgca tatctcatta aagcaggaca agctcaaacc tttgcaaatg cgttttcaag
     2101 caaagaagcg attttctgag ttgatgggtt gttccttgca gccatgatca tcttagcaag
     2161 tgcgtcctca gtgaaaagtg cctggaagat tttagccata gattgtttct ggtctgggta
     2221 ctttgtattc caatcatcca aatacttaga ccaaagctga aactctggag atgaaagaag
     2281 tccgtctgac accttattca atctaacttg gtacaacttt ccgtccaacc tctcgttgta
     2341 cctgataatc acatcagctc ctggtttttc aaggatgaat gtgttgaatg cgtccctagg
     2401 tggcctctca gcatgcaacc acctcttgta tctctggatc tgaagctgtt ttgacaattc
     2461 aacagtagat ggattcttca taccctcagc cacgatctta gccactccgt ggtcgtggta
     2521 gtacctgtgg ataggagcca acaaagtctc cttatcagtg tttttcttgt taaatgactc
     2581 caaataattg aaccatgtct tgaacattgg tgttgacaaa aggttatcac cagtctcatg
     2641 aagattgaaa gcttcaaaca catactctgg agatctgtta gagttaagaa gccttttgat
     2701 ctgctcagcc tgcaaccttg aagccacgtt ctctgtaagc tgcacctttt tagctgcctc
     2761 caacatagcg aacactgcgt cgtcaccgta gtgagcagca attggaggct ccattgaagc
     2821 cctagatgtt gggtactgaa ggttgaagtc atccacgtac tttgtccaag cgataaacct
     2881 agggtttgtc aacaatccgt ccaccttgtc gtcgatcatg aacatctcaa acacatattt
     2941 tggtgtctgt gcgtcgtcca accatctttg gatctgctct ttcctcaact tagtagccaa
     3001 tgcgtttgta gtagagtatg tcttagccaa ctcaagagct cttgtcaaac cctcgtcaga
     3061 atagatagtc ctcaatgttg ggatcataga ctctttctta gttgggtttt ttgcgttaaa
     3121 gtcgtcaacg taattaaccc atgtattgaa gtcaggcaat ccaaacacag ttccctcgat
     3181 cttcttgaag ttaggcacca tgaacttgtt caattccaac aactggaaca cttcctgagt
     3241 tgactttccc tcgtccaacc acttctgagt ttgcaaagtc ttaagcctac cagctgcctc
     3301 aactgccttg ttcacaggct ttgcagcacc caaaagctga gacaactttt cgtctggcac
     3361 gtccctctgt gatggcttag actttgttgt gatcttgtta gtcaatgact gaaacatagc
     3421 ttcaatcttt tcaagtgatg gtcctgatgg gaatcccatt gtatcgataa ttgtaaatgt
     3481 aattgtaatg ttgtttgttg tttgttgttg ttggtaattg ttgtaaaaat gagctcttat
     3541 actcgagcgt gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga
     3601 aggatagtgg gattgtgcgt catcccttac gtcagtggag atgtcacatc aatccacttg
     3661 ctttgtagac gtggttggaa cctcttcttt ttccacgatg ctcctcgtgg gtgggggtcc
     3721 atctttggga ccactgtcgg cagagagatc ttgaatgata gcctttcctt tatcgcaatg
     3781 atggcatttg taggagccac cttccttttc tactgtcctt tcgatgaagt gacagatagc
     3841 tgggcaatgg aatccgagga ggtttcccga aattatcctt tgttgaaaag tctcaatagc
     3901 cctttgatct tctgagactg tatctttgac atttttggag tagaccagag tgtcgtgctc
     3961 caccatgttg acgaagattt tcttcttgtc attgagtcgt aaaagactct gtatgaactg
     4021 ttcgccagtc ttgacggcga gttctgttag atcctcgatt tgaatcttag actccatgca
     4081 tggccttaga ttcagtagga actacctttt tagagactcc aatctctatt acttgccttg
     4141 gtttatgaag caagccttga atcgtccata ctggaatagt acttctgatc ttgagaaata
     4201 tgtctttctc tgtgttcttg atgcaattag tcctgaatct tttgactgca tctttaacct
     4261 tcttgggaag gtatttgatc tcctggagat tgttactcgg gtagatcgtc ttgatgagac
     4321 ttgctgcgta ggcctctcta accatctgtg ggtcagcatt ctttctgaaa ttgaagaggc
     4381 taaccttctc attatcagtg gtgaacatag tgtcgtcacc ttcaccttcg aacttccttc
     4441 ctagatcgta aagatagagg aaatcgtcca ttgtaatctc cggggcaaag gagatctctt
     4501 ttggggctgg atcactgctg ggccttttgg ttcctagcgt gagccagtgg gctttttgct
     4561 ttggtgggct tgttagggcc ttagcaaagc tcttgggctt gagttgagct tctcctttgg
     4621 ggatgaagtt caacctgtct gtttgctgac ttgttgtgta cgcgtcagct gctgctcttg
     4681 cctctgtaat agtggcaaat ttcttgtgtg caactccggg aacgccgttt gttgccgcct
     4741 ttgtacaacc ccagtcatcg tatataccgg catgtggacc gttatacaca acgtagtagt
     4801 tgatatgagg gtgttgaata cccgattctg ctctgagagg agcaactgtg ctgttaagct
     4861 cagatttttg tgggattgga attcctccaa gcttgaattc ttgcagcgct ccaattcagc
     4921 aagcgtcgat ctagtaacat agatgacacc gcgcgcgata atttatccta gtttgcgcgc
     4981 tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat cataaaaacc
     5041 catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac gtaattcaac
     5101 agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa gaaactttat
     5161 tgccaaatgt ttgaacgatc tgcttgactc tagatccaga gtcccgctca aatctcagtc
     5221 acaggaagaa caggtctagg aggaacagga agagagaaat caagctgcca gaatcccaca
     5281 tcatgccaat ttccatgttt gaaaccagca gctctaagca ttcccctagg agcataacca
     5341 agagcctcat gcatcctcac agaaggatca ttaggaagtc caatcacagc aaccacagac
     5401 ttgaaaccct gagcctccaa agacttcaaa agatgagtgt aaagagtaga tccaagacca
     5461 gttctttgat gcctaggaga cacgtaaaca gtagactcag cagtccaatc gtaagcattc
     5521 ctagccttcc aaggaccagc ataagcaata ccagccactt caccatccac ttcagcaaca
     5581 agccaaggat acctctctct aagcctcaca agatcatcag tccactcttg tggttcttga
     5641 ggctcagtcc tgaaattcac agtagaagtc tcaatgtaat gattcacaat agtgcacaca
     5701 gcaggcatat cagcctcagt agcccttcta atatcagcag gccttctttc aggagacatg
     5761 cgaaacgatc cagatccggt gcagattatt tggattgaga gtgaatatga gactctaatt
     5821 ggataccgag gggaatttat ggaacgtcag tggagcattt tgacaagaaa tatttgctag
     5881 ctgatagtga ccttaggcga cttttgaacg cgcaataatg gtttctgacg tatgtgctta
     5941 gctcattaaa ctccagaaac ccgcggctga gtggctcctt caacgttgcg gttctgtcag
     6001 ttccaaacgt aaaacggctt gtcccgcgtc atcggcgggg gtcataacgt gactccctta
     6061 attctccgct ccggatccga attcggcaca gagtgttcaa ccccagtaca ttaaaaacgt
     6121 ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc
     6181 accagccagc caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg
     6241 cagcccatca accatgacta ctgtccggtc caacagatcc gagcaggaac cgtaaaaagg
     6301 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6361 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6421 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6481 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6541 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6601 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6661 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6721 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6781 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6841 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6901 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6961 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     7021 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagctaaa
     7081 acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg atccccagta
     7141 agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgagc gaccggacgc
     7201 agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca ataaagccac
     7261 ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg gaaaagacaa
     7321 gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg
     7381 gagtgtcctc ttcccagttt tcgcaatcca catcggccag atcgttattc agtaagtaat
     7441 ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat atgtcgatgg
     7501 agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg ctttgttcat
     7561 cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga ttgctccagc
     7621 catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc agccatagca
     7681 tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg tccgtcattt
     7741 ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga gacattcctt
     7801 ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt gatattctca
     7861 ttttagccat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7921 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcactc
     7981 ggagccttga tgtgggcgct ttttgcagct gttcggctgt gcgctggcca gacagttatg
     8041 cacaggccag gcgggtttta agagttttaa taagttttaa agagttttag gcggaaaaat
     8101 cgcctttttt ctcttttata tcagtcactt acatgtgtga ccggttccca atgtacggct
     8161 ttgggttccc aatgtacggg ttccggttcc caatgtacgg ctttgggttc ccaatgtacg
     8221 tgctatccac aggaaagaag ctttttcgac ctttttcccc tgctagggca atttgcccta
     8281 gcatctgctc cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg ttgcggtagc
     8341 gcatgactag gatcgggcca gcctgccccg cctcctcctt caaatcgtac tccggcaggt
     8401 catttgaccc gatcagcttg cgcacggtga aacagaactt cttgaactct ccggcgctgc
     8461 cactgcgttc gtagatcgtc ttgaacaacc atctggcttc tgccttgcct gcggcgcggc
     8521 gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa tcggggtgaa
     8581 ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg gtacatccaa tcagctagct
     8641 cgatctcgat gtactccggc cgcccggttt cgctctttac gatcttgtag cggctaatca
     8701 aggcttcacc ctcggatacc gtcaccaggc ggccgttctt ggccttcttc gtacgctgca
     8761 tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac caggtcgtct ttctgctttc
     8821 cgccatcggc tcgccggcag aacttgagta cgtccgcaac gtgtggacgg aacacgcggc
     8881 cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga tgggaaaccg
     8941 ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct gcggaaacct
     9001 ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg tcacgcttcg
     9061 acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc acgtcataga
     9121 gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta atcgacgacg
     9181 caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc gcttgccacg
     9241 attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc gcggccttca
     9301 acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg gatggtttgc
     9361 gaccgctcac gccgattcct cgggcttggg ggttccagtg ccattgcagg gccggcagac
     9421 aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatgggg
//
