LOCUS       sequence-342205-        8413 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 8413)
  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 8413)
  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_10232
FEATURES             Location/Qualifiers
     source          1..8413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    640..664
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(731..1438)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1866..2211)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1903..1923)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3244..3496)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4096..4278)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4132..4310)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4473..4497
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4633..5221)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4713..4732)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5392..6186)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6187..6291)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6353..6547
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6613..7686)
                     /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"
     CDS             complement(join(8115..8413,1..331))
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
ORIGIN
        1 cgtaagggct tggctgcacc ggaatcagca cgaagtcggc tgccttgatc gcggacacag
       61 ccaagtccgc cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg ccgatggcct
      121 tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt tgatcttccc
      181 gcacggccgc ccaatcgcgg gcactgccct ggggatcgga atcgactaac agaacatcgg
      241 ccccggcgag ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg cctgacccgc
      301 ctttctggtt aagtacagcg ataaccttca tgcgttcccc ttgcgtattt gtttatttac
      361 tcatcgcatc atatacgcag cgaccgcatg acgcaagctg ttttactcaa atacacatca
      421 cctttttaga cggcggcgct cggtttcttc agcggccaag ctggccggcc aggccgccag
      481 cttggcatca gacaaaccgg ccaggatttc atgcagccgc acggctgact cccgagggga
      541 accctgtggt tggcatgcac atacaaatgg acgaacggat aaaccttttc acgccctttt
      601 aaatatccga ttattctaat aaacgctctt ttctcttagg tttacccgcc aatatatcct
      661 gtcaaacact gatagtttaa acggatcact tcgtgtccct cggtcacatg tgcatcctct
      721 agtagcggtc ctgctgagcc tcgacatgtt gtcgcaaaat tcgccctgga cccgcccaac
      781 gatttgtcgt cactgtcaag gtttgacctg cacttcattt ggggcccaca tacaccaaaa
      841 aaatgctgca taattctcgg ggcagcaagt cggttacccg gccgccgtgc tggaccgggt
      901 tgaatggtgc ccgtaacttt cggtagagcg gacggccaat actcaacttc aaggaatctc
      961 acccatgcgc gccggcgggg aaccggagtt cccttcagtg agcgttatta gttcgccgct
     1021 cggtgtgtcg tagatactag cccctggggc acttttgaaa tttgaataag atttatgtaa
     1081 tcagtctttt aggtttgacc ggttctgccg ctttttttaa aattggattt gtaataataa
     1141 aacgcaattg tttgttattg tggcgctcta tcatagatgt cgctataaac ctattcagca
     1201 caatatattg ttttcatttt aatattgtac atataagtag tagggtacaa tcagtaaatt
     1261 gaacggagaa tattattcat aaaaatacga tagtaacggg tgatatattc attagaatga
     1321 accgaaaccg gcggtaagga tctgagctac acatgctcag gttttttaca acgtgcacaa
     1381 cagaattgaa agcaaatatc atgcgatcat aggcttctcg catatctcat taaagcagga
     1441 caagctcaag tagtagaagc caccctctgt ccgttcctct gcataccgtc gtagaatcca
     1501 gcgtactgct tgaacctcct gtaagcctga gacttccaga tgtagtctga tgggaactgg
     1561 tgctggtgca ccatagtcac ccactgagaa agagacatct ccacagactc gtccttgtgc
     1621 ctgatcacgt cgaacaacct gtgaggctgc actccagcag cgtacaactc agcgtcagta
     1681 gcgtccatct tgatcaagtt agcaatttgg ttcttcatct tcctctccct aagagggtgc
     1741 atagcagcgt tagtcaacac agtcaagtat cccattgtat cgataattgt aaatgtaatt
     1801 gtaatgttgt ttgttgtttg ttgttgttgg taattgttgt aaaaatgagc tcttatactc
     1861 gagcgtgtcc tctccaaatg aaatgaactt ccttatatag aggaagggtc ttgcgaagga
     1921 tagtgggatt gtgcgtcatc ccttacgtca gtggagatgt cacatcaatc cacttgcttt
     1981 gtagacgtgg ttggaacctc ttctttttcc acgatgctcc tcgtgggtgg gggtccatct
     2041 ttgggaccac tgtcggcaga gagatcttga atgatagcct ttcctttatc gcaatgatgg
     2101 catttgtagg agccaccttc cttttctact gtcctttcga tgaagtgaca gatagctggg
     2161 caatggaatc cgaggaggtt tcccgaaatt atcctttgtt gaaaagtctc aatagccctt
     2221 tgatcttctg agactgtatc tttgacattt ttggagtaga ccagagtgtc gtgctccacc
     2281 atgttgacga agattttctt cttgtcattg agtcgtaaaa gactctgtat gaactgttcg
     2341 ccagtcttga cggcgagttc tgttagatcc tcgatttgaa tcttagactc catgcatggc
     2401 cttagattca gtaggaacta cctttttaga gactccaatc tctattactt gccttggttt
     2461 atgaagcaag ccttgaatcg tccatactgg aatagtactt ctgatcttga gaaatatgtc
     2521 tttctctgtg ttcttgatgc aattagtcct gaatcttttg actgcatctt taaccttctt
     2581 gggaaggtat ttgatctcct ggagattgtt actcgggtag atcgtcttga tgagacttgc
     2641 tgcgtaggcc tctctaacca tctgtgggtc agcattcttt ctgaaattga agaggctaac
     2701 cttctcatta tcagtggtga acatagtgtc gtcaccttca ccttcgaact tccttcctag
     2761 atcgtaaaga tagaggaaat cgtccattgt aatctccggg gcaaaggaga tctcttttgg
     2821 ggctggatca ctgctgggcc ttttggttcc tagcgtgagc cagtgggctt tttgctttgg
     2881 tgggcttgtt agggccttag caaagctctt gggcttgagt tgagcttctc ctttggggat
     2941 gaagttcaac ctgtctgttt gctgacttgt tgtgtacgcg tcagctgctg ctcttgcctc
     3001 tgtaatagtg gcaaatttct tgtgtgcaac tccgggaacg ccgtttgttg ccgcctttgt
     3061 acaaccccag tcatcgtata taccggcatg tggaccgtta tacacaacgt agtagttgat
     3121 atgagggtgt tgaatacccg attctgctct gagaggagca actgtgctgt taagctcaga
     3181 tttttgtggg attggaattc ctccaagctt gaattcttgc agcgctccaa ttcagcaagc
     3241 gtcgatctag taacatagat gacaccgcgc gcgataattt atcctagttt gcgcgctata
     3301 ttttgttttc tatcgcgtat taaatgtata attgcgggac tctaatcata aaaacccatc
     3361 tcataaataa cgtcatgcat tacatgttaa ttattacatg cttaacgtaa ttcaacagaa
     3421 attatatgat aatcatcgca agaccggcaa caggattcaa tcttaagaaa ctttattgcc
     3481 aaatgtttga acgatctgct tgactctaga tccagagtcc cgctcaaatc tcagtcacag
     3541 gaagaacagg tctaggagga acaggaagag agaaatcaag ctgccagaat cccacatcat
     3601 gccaatttcc atgtttgaaa ccagcagctc taagcattcc cctaggagca taaccaagag
     3661 cctcatgcat cctcacagaa ggatcattag gaagtccaat cacagcaacc acagacttga
     3721 aaccctgagc ctccaaagac ttcaaaagat gagtgtaaag agtagatcca agaccagttc
     3781 tttgatgcct aggagacacg taaacagtag actcagcagt ccaatcgtaa gcattcctag
     3841 ccttccaagg accagcataa gcaataccag ccacttcacc atccacttca gcaacaagcc
     3901 aaggatacct ctctctaagc ctcacaagat catcagtcca ctcttgtggt tcttgaggct
     3961 cagtcctgaa attcacagta gaagtctcaa tgtaatgatt cacaatagtg cacacagcag
     4021 gcatatcagc ctcagtagcc cttctaatat cagcaggcct tctttcagga gacatgcgaa
     4081 acgatccaga tccggtgcag attatttgga ttgagagtga atatgagact ctaattggat
     4141 accgagggga atttatggaa cgtcagtgga gcattttgac aagaaatatt tgctagctga
     4201 tagtgacctt aggcgacttt tgaacgcgca ataatggttt ctgacgtatg tgcttagctc
     4261 attaaactcc agaaacccgc ggctgagtgg ctccttcaac gttgcggttc tgtcagttcc
     4321 aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact cccttaattc
     4381 tccgctccgg atccgaattc ggcacagagt gttcaacccc agtacattaa aaacgtccgc
     4441 aatgtgttat taagttgtct aagcgtcaat ttgtttacac cacaatatat cctgccacca
     4501 gccagccaac agctccccga ccggcagctc ggcacaaaat caccactcga tacaggcagc
     4561 ccatcaacca tgactactgt ccggtccaac agatccgagc aggaaccgta aaaaggccgc
     4621 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4681 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4741 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4801 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     4861 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4921 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4981 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5041 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     5101 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5161 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5221 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5281 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5341 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gctaaaacaa
     5401 ttcatccagt aaaatataat attttatttt ctcccaatca ggcttgatcc ccagtaagtc
     5461 aaaaaatagc tcgacatact gttcttcccc gatatcctcc ctgagcgacc ggacgcagaa
     5521 ggcaatgtca taccacttgt ccgccctgcc gcttctccca agatcaataa agccacttac
     5581 tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg ccgtgggaaa agacaagttc
     5641 ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg cgcggatctt taaatggagt
     5701 gtcctcttcc cagttttcgc aatccacatc ggccagatcg ttattcagta agtaatccaa
     5761 ttcggctaag cggctgtcta agctattcgt atagggacaa tccgatatgt cgatggagtg
     5821 aaagagcctg atgcactccg catacagctc gataatcttt tcagggcttt gttcatcttc
     5881 atactcttcc gagcaaagga cgccatcggc ctcactcatg agcagattgc tccagccatc
     5941 atgccgttca aagtgcagga cctttggaac aggcagcttt ccttccagcc atagcatcat
     6001 gtccttttcc cgttccacat cataggtggt ccctttatac cggctgtccg tcatttttaa
     6061 atataggttt tcattttctc ccaccagctt atatacctta gcaggagaca ttccttccgt
     6121 atcttttacg cagcggtatt tttcgatcag ttttttcaat tccggtgata ttctcatttt
     6181 agccatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6241 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcactcggag
     6301 ccttgatgtg ggcgcttttt gcagctgttc ggctgtgcgc tggccagaca gttatgcaca
     6361 ggccaggcgg gttttaagag ttttaataag ttttaaagag ttttaggcgg aaaaatcgcc
     6421 ttttttctct tttatatcag tcacttacat gtgtgaccgg ttcccaatgt acggctttgg
     6481 gttcccaatg tacgggttcc ggttcccaat gtacggcttt gggttcccaa tgtacgtgct
     6541 atccacagga aagaagcttt ttcgaccttt ttcccctgct agggcaattt gccctagcat
     6601 ctgctccgta cattaggaac cggcggatgc ttcgccctcg atcaggttgc ggtagcgcat
     6661 gactaggatc gggccagcct gccccgcctc ctccttcaaa tcgtactccg gcaggtcatt
     6721 tgacccgatc agcttgcgca cggtgaaaca gaacttcttg aactctccgg cgctgccact
     6781 gcgttcgtag atcgtcttga acaaccatct ggcttctgcc ttgcctgcgg cgcggcgtgc
     6841 caggcggtag agaaaacggc cgatgccggg atcgatcaaa aagtaatcgg ggtgaaccgt
     6901 cagcacgtcc gggttcttgc cttctgtgat ctcgcggtac atccaatcag ctagctcgat
     6961 ctcgatgtac tccggccgcc cggtttcgct ctttacgatc ttgtagcggc taatcaaggc
     7021 ttcaccctcg gataccgtca ccaggcggcc gttcttggcc ttcttcgtac gctgcatggc
     7081 aacgtgcgtg gtgtttaacc gaatgcaggt ttctaccagg tcgtctttct gctttccgcc
     7141 atcggctcgc cggcagaact tgagtacgtc cgcaacgtgt ggacggaaca cgcggccggg
     7201 cttgtctccc ttcccttccc ggtatcggtt catggattcg gttagatggg aaaccgccat
     7261 cagtaccagg tcgtaatccc acacactggc catgccggcc ggccctgcgg aaacctctac
     7321 gtgcccgtct ggaagctcgt agcggatcac ctcgccagct cgtcggtcac gcttcgacag
     7381 acggaaaacg gccacgtcca tgatgctgcg actatcgcgg gtgcccacgt catagagcat
     7441 cggaacgaaa aaatctggtt gctcgtcgcc cttgggcggc ttcctaatcg acgacgcacc
     7501 ggctgccggc ggttgccggg attctttgcg gattcgatca gcggccgctt gccacgattc
     7561 accggggcgt gcttctgcct cgatgcgttg ccgctgggcg gcctgcgcgg ccttcaactt
     7621 ctccaccagg tcatcaccca gcgccgcgcc gatttgtacc gggccggatg gtttgcgacc
     7681 gctcacgccg attcctcggg cttgggggtt ccagtgccat tgcagggccg gcagacaacc
     7741 cagccgctta cgcctggcca accgcccgtt cctccacaca tggggcattc cacggcgtcg
     7801 gtgcctggtt gttcttgatt ttccatgccg cctcctttag ccgctaaaat tcatctactc
     7861 atttattcat ttgctcattt actctggtag ctgcgcgatg tattcagata gcagctcggt
     7921 aatggtcttg ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct ccgccggcaa
     7981 ctgaaagttg acccgcttca tggctggcgt gtctgccagg ctggccaacg ttgcagcctt
     8041 gctgctgcgt gcgctcggac ggccggcact tagcgtgttt gtgcttttgc tcattttctc
     8101 tttacctcat taactcaaat gagttttgat ttaatttcag cggccagcgc ctggacctcg
     8161 cgggcagcgt cgccctcggg ttctgattca agaacggttg tgccggcggc ggcagtgcct
     8221 gggtagctca cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta cccggccagc
     8281 gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac gacaaaggcc
     8341 gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa ccagctccac caggtcggcg
     8401 gtggcccata tgt
//
