LOCUS       sequence-342791-        9364 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 9364)
  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 9364)
  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_20922
FEATURES             Location/Qualifiers
     source          1..9364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(176..805)
                     /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    1114..1138
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1205..1912)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3291..3636)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3328..3348)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4669..4921)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5521..5703)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5557..5735)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5898..5922
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6058..6646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6138..6157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6817..7611)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7612..7716)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7778..7972
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8038..9111)
                     /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 accgcgtaca tcttcagctt ggtgtgatcc tccgccggca actgaaagtt gacccgcttc
       61 atggctggcg tgtctgccag gctggccaac gttgcagcct tgctgctgcg tgcgctcgga
      121 cggccggcac ttagcgtgtt tgtgcttttg ctcattttct ctttacctca ttaactcaaa
      181 tgagttttga tttaatttca gcggccagcg cctggacctc gcgggcagcg tcgccctcgg
      241 gttctgattc aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg
      301 tgatacggga ctcaagaatg ggcagctcgt acccggccag cgcctcggca acctcaccgc
      361 cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg
      421 tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag
      481 ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt
      541 ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt
      601 cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg
      661 ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg
      721 cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct
      781 ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg
      841 catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt
      901 tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc
      961 atcagacaaa ccggccagga tttcatgcag ccgcacggct gactcccgag gggaaccctg
     1021 tggttggcat gcacatacaa atggacgaac ggataaacct tttcacgccc ttttaaatat
     1081 ccgattattc taataaacgc tcttttctct taggtttacc cgccaatata tcctgtcaaa
     1141 cactgatagt ttaaacggat cacttcgtgt ccctcggtca catgtgcatc ctctagtagc
     1201 ggtcctgctg agcctcgaca tgttgtcgca aaattcgccc tggacccgcc caacgatttg
     1261 tcgtcactgt caaggtttga cctgcacttc atttggggcc cacatacacc aaaaaaatgc
     1321 tgcataattc tcggggcagc aagtcggtta cccggccgcc gtgctggacc gggttgaatg
     1381 gtgcccgtaa ctttcggtag agcggacggc caatactcaa cttcaaggaa tctcacccat
     1441 gcgcgccggc ggggaaccgg agttcccttc agtgagcgtt attagttcgc cgctcggtgt
     1501 gtcgtagata ctagcccctg gggcactttt gaaatttgaa taagatttat gtaatcagtc
     1561 ttttaggttt gaccggttct gccgcttttt ttaaaattgg atttgtaata ataaaacgca
     1621 attgtttgtt attgtggcgc tctatcatag atgtcgctat aaacctattc agcacaatat
     1681 attgttttca ttttaatatt gtacatataa gtagtagggt acaatcagta aattgaacgg
     1741 agaatattat tcataaaaat acgatagtaa cgggtgatat attcattaga atgaaccgaa
     1801 accggcggta aggatctgag ctacacatgc tcaggttttt tacaacgtgc acaacagaat
     1861 tgaaagcaaa tatcatgcga tcataggctt ctcgcatatc tcattaaagc aggacaagct
     1921 cactgcctag tagcgtaaaa gaatgtataa aaagacctgt acacatccca gtactttggg
     1981 tcagcgtacc tcaactcctt cacattccac ctcttctcct ggatccactg tttgaactga
     2041 agattctgca atgatgtgat tgtctgagct gtgttaaggt cagggttctt tttagccaca
     2101 gcgcaaagag cccttgccaa gtcaaggttg ttgtcaaagt gctttgaaag ctcagcgatg
     2161 atagcgtact cgtcctggtt cattttcttc ctctgaagct tgttgaagta agacacccag
     2221 attccgaaag cagggtcaga caaagcgttg tcacccttac cagacaacct caacaacttg
     2281 aaagcttcat gtgctggcat tccgtccctc aaccaaagtg ctgatgccac ctgagcagca
     2341 gtagccttca agtttccagt gatagaagaa tgtgtcacca ttttcttaag tgtctcatca
     2401 tcgtaatatt tagtcatcac agatgacatc acgttgtcag cctcctcttt gtgcatagat
     2461 ttcacatatg tgaaccaagt aaccaagatt ggattttcga agaaattaat accgtccttg
     2521 tcaagagaca agtgagtgaa cacgtcctct gcagtctttt tctgtgacaa ccagtttccc
     2581 ttgataagcc tcaccatagg tcccaatgtg aacttcaatg tgtcgtcgtt ctttccagcg
     2641 aaaaacattt tcaagatctc tgcgtctgaa aacctaatct caagtttcct gtaaagcacc
     2701 tcagttgggt tctcagagat ctttttcacg taaaggaacc aagacaccaa acctggtgac
     2761 ttcaaagtat caatagattg gtcgatgtcc aacctaagca ttttgaacac atcatcagat
     2821 gtctttccct tttccttcca aatcttgaac tgtgtagatt caagcttcac agccattgac
     2881 acaaggtcag agtcagactt tgccctcacc aacatctttg gcaactcctc ctcaccgtgc
     2941 ctctttgcca acttcaaaac caagaattcg tatgggtcca atgtatgatg tttagtagcg
     3001 taagaaatga atgtttgaag agctgggtgc ttaagaagtc caagctcctt gtcagcaggc
     3061 accaatgaga tcttaagcac gtcgtcctct gcctttcctg tagtcaacca cttcttgatc
     3121 tgagcgaact caagtccaga agcgatgtcc ctagtagaag ctgtagtacc agctgccatc
     3181 aacctcttgt gcaaagccat tgtatcgata attgtaaatg taattgtaat gttgtttgtt
     3241 gtttgttgtt gttggtaatt gttgtaaaaa tgagctctta tactcgagcg tgtcctctcc
     3301 aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg
     3361 tcatccctta cgtcagtgga gatgtcacat caatccactt gctttgtaga cgtggttgga
     3421 acctcttctt tttccacgat gctcctcgtg ggtgggggtc catctttggg accactgtcg
     3481 gcagagagat cttgaatgat agcctttcct ttatcgcaat gatggcattt gtaggagcca
     3541 ccttcctttt ctactgtcct ttcgatgaag tgacagatag ctgggcaatg gaatccgagg
     3601 aggtttcccg aaattatcct ttgttgaaaa gtctcaatag ccctttgatc ttctgagact
     3661 gtatctttga catttttgga gtagaccaga gtgtcgtgct ccaccatgtt gacgaagatt
     3721 ttcttcttgt cattgagtcg taaaagactc tgtatgaact gttcgccagt cttgacggcg
     3781 agttctgtta gatcctcgat ttgaatctta gactccatgc atggccttag attcagtagg
     3841 aactaccttt ttagagactc caatctctat tacttgcctt ggtttatgaa gcaagccttg
     3901 aatcgtccat actggaatag tacttctgat cttgagaaat atgtctttct ctgtgttctt
     3961 gatgcaatta gtcctgaatc ttttgactgc atctttaacc ttcttgggaa ggtatttgat
     4021 ctcctggaga ttgttactcg ggtagatcgt cttgatgaga cttgctgcgt aggcctctct
     4081 aaccatctgt gggtcagcat tctttctgaa attgaagagg ctaaccttct cattatcagt
     4141 ggtgaacata gtgtcgtcac cttcaccttc gaacttcctt cctagatcgt aaagatagag
     4201 gaaatcgtcc attgtaatct ccggggcaaa ggagatctct tttggggctg gatcactgct
     4261 gggccttttg gttcctagcg tgagccagtg ggctttttgc tttggtgggc ttgttagggc
     4321 cttagcaaag ctcttgggct tgagttgagc ttctcctttg gggatgaagt tcaacctgtc
     4381 tgtttgctga cttgttgtgt acgcgtcagc tgctgctctt gcctctgtaa tagtggcaaa
     4441 tttcttgtgt gcaactccgg gaacgccgtt tgttgccgcc tttgtacaac cccagtcatc
     4501 gtatataccg gcatgtggac cgttatacac aacgtagtag ttgatatgag ggtgttgaat
     4561 acccgattct gctctgagag gagcaactgt gctgttaagc tcagattttt gtgggattgg
     4621 aattcctcca agcttgaatt cttgcagcgc tccaattcag caagcgtcga tctagtaaca
     4681 tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg ttttctatcg
     4741 cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata aataacgtca
     4801 tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat atgataatca
     4861 tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg tttgaacgat
     4921 ctgcttgact ctagatccag agtcccgctc aaatctcagt cacaggaaga acaggtctag
     4981 gaggaacagg aagagagaaa tcaagctgcc agaatcccac atcatgccaa tttccatgtt
     5041 tgaaaccagc agctctaagc attcccctag gagcataacc aagagcctca tgcatcctca
     5101 cagaaggatc attaggaagt ccaatcacag caaccacaga cttgaaaccc tgagcctcca
     5161 aagacttcaa aagatgagtg taaagagtag atccaagacc agttctttga tgcctaggag
     5221 acacgtaaac agtagactca gcagtccaat cgtaagcatt cctagccttc caaggaccag
     5281 cataagcaat accagccact tcaccatcca cttcagcaac aagccaagga tacctctctc
     5341 taagcctcac aagatcatca gtccactctt gtggttcttg aggctcagtc ctgaaattca
     5401 cagtagaagt ctcaatgtaa tgattcacaa tagtgcacac agcaggcata tcagcctcag
     5461 tagcccttct aatatcagca ggccttcttt caggagacat gcgaaacgat ccagatccgg
     5521 tgcagattat ttggattgag agtgaatatg agactctaat tggataccga ggggaattta
     5581 tggaacgtca gtggagcatt ttgacaagaa atatttgcta gctgatagtg accttaggcg
     5641 acttttgaac gcgcaataat ggtttctgac gtatgtgctt agctcattaa actccagaaa
     5701 cccgcggctg agtggctcct tcaacgttgc ggttctgtca gttccaaacg taaaacggct
     5761 tgtcccgcgt catcggcggg ggtcataacg tgactccctt aattctccgc tccggatccg
     5821 aattcggcac agagtgttca accccagtac attaaaaacg tccgcaatgt gttattaagt
     5881 tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc caccagccag ccaacagctc
     5941 cccgaccggc agctcggcac aaaatcacca ctcgatacag gcagcccatc aaccatgact
     6001 actgtccggt ccaacagatc cgagcaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6481 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6781 aatctaaagt atatatgagt aaacttggtc tgacagctaa aacaattcat ccagtaaaat
     6841 ataatatttt attttctccc aatcaggctt gatccccagt aagtcaaaaa atagctcgac
     6901 atactgttct tccccgatat cctccctgag cgaccggacg cagaaggcaa tgtcatacca
     6961 cttgtccgcc ctgccgcttc tcccaagatc aataaagcca cttactttgc catctttcac
     7021 aaagatgttg ctgtctccca ggtcgccgtg ggaaaagaca agttcctctt cgggcttttc
     7081 cgtctttaaa aaatcataca gctcgcgcgg atctttaaat ggagtgtcct cttcccagtt
     7141 ttcgcaatcc acatcggcca gatcgttatt cagtaagtaa tccaattcgg ctaagcggct
     7201 gtctaagcta ttcgtatagg gacaatccga tatgtcgatg gagtgaaaga gcctgatgca
     7261 ctccgcatac agctcgataa tcttttcagg gctttgttca tcttcatact cttccgagca
     7321 aaggacgcca tcggcctcac tcatgagcag attgctccag ccatcatgcc gttcaaagtg
     7381 caggaccttt ggaacaggca gctttccttc cagccatagc atcatgtcct tttcccgttc
     7441 cacatcatag gtggtccctt tataccggct gtccgtcatt tttaaatata ggttttcatt
     7501 ttctcccacc agcttatata ccttagcagg agacattcct tccgtatctt ttacgcagcg
     7561 gtatttttcg atcagttttt tcaattccgg tgatattctc attttagcca tactcttcct
     7621 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7681 atgtatttag aaaaataaac aaataggggt tccgcgcact cggagccttg atgtgggcgc
     7741 tttttgcagc tgttcggctg tgcgctggcc agacagttat gcacaggcca ggcgggtttt
     7801 aagagtttta ataagtttta aagagtttta ggcggaaaaa tcgccttttt tctcttttat
     7861 atcagtcact tacatgtgtg accggttccc aatgtacggc tttgggttcc caatgtacgg
     7921 gttccggttc ccaatgtacg gctttgggtt cccaatgtac gtgctatcca caggaaagaa
     7981 gctttttcga cctttttccc ctgctagggc aatttgccct agcatctgct ccgtacatta
     8041 ggaaccggcg gatgcttcgc cctcgatcag gttgcggtag cgcatgacta ggatcgggcc
     8101 agcctgcccc gcctcctcct tcaaatcgta ctccggcagg tcatttgacc cgatcagctt
     8161 gcgcacggtg aaacagaact tcttgaactc tccggcgctg ccactgcgtt cgtagatcgt
     8221 cttgaacaac catctggctt ctgccttgcc tgcggcgcgg cgtgccaggc ggtagagaaa
     8281 acggccgatg ccgggatcga tcaaaaagta atcggggtga accgtcagca cgtccgggtt
     8341 cttgccttct gtgatctcgc ggtacatcca atcagctagc tcgatctcga tgtactccgg
     8401 ccgcccggtt tcgctcttta cgatcttgta gcggctaatc aaggcttcac cctcggatac
     8461 cgtcaccagg cggccgttct tggccttctt cgtacgctgc atggcaacgt gcgtggtgtt
     8521 taaccgaatg caggtttcta ccaggtcgtc tttctgcttt ccgccatcgg ctcgccggca
     8581 gaacttgagt acgtccgcaa cgtgtggacg gaacacgcgg ccgggcttgt ctcccttccc
     8641 ttcccggtat cggttcatgg attcggttag atgggaaacc gccatcagta ccaggtcgta
     8701 atcccacaca ctggccatgc cggccggccc tgcggaaacc tctacgtgcc cgtctggaag
     8761 ctcgtagcgg atcacctcgc cagctcgtcg gtcacgcttc gacagacgga aaacggccac
     8821 gtccatgatg ctgcgactat cgcgggtgcc cacgtcatag agcatcggaa cgaaaaaatc
     8881 tggttgctcg tcgcccttgg gcggcttcct aatcgacgac gcaccggctg ccggcggttg
     8941 ccgggattct ttgcggattc gatcagcggc cgcttgccac gattcaccgg ggcgtgcttc
     9001 tgcctcgatg cgttgccgct gggcggcctg cgcggccttc aacttctcca ccaggtcatc
     9061 acccagcgcc gcgccgattt gtaccgggcc ggatggtttg cgaccgctca cgccgattcc
     9121 tcgggcttgg gggttccagt gccattgcag ggccggcaga caacccagcc gcttacgcct
     9181 ggccaaccgc ccgttcctcc acacatgggg cattccacgg cgtcggtgcc tggttgttct
     9241 tgattttcca tgccgcctcc tttagccgct aaaattcatc tactcattta ttcatttgct
     9301 catttactct ggtagctgcg cgatgtattc agatagcagc tcggtaatgg tcttgccttg
     9361 gcgt
//
