LOCUS       sequence-342207-        8356 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 8356)
  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 8356)
  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_10654
FEATURES             Location/Qualifiers
     source          1..8356
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    734..758
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(825..1532)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1903..2248)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1940..1960)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3281..3533)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4133..4315)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4169..4347)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4510..4534
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4670..5258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4750..4769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5429..6223)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6224..6328)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6390..6584
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6650..7723)
                     /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(8152..8356,1..425))
                     /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 gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg tgacctcaat gcgctgctta
       61 accagctcca ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg caccggaatc
      121 agcacgaagt cggctgcctt gatcgcggac acagccaagt ccgccgcctg gggcgctccg
      181 tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat cgtcgggcgg
      241 tcgatgccga caacggttag cggttgatct tcccgcacgg ccgcccaatc gcgggcactg
      301 ccctggggat cggaatcgac taacagaaca tcggccccgg cgagttgcag ggcgcgggct
      361 agatgggttg cgatggtcgt cttgcctgac ccgcctttct ggttaagtac agcgataacc
      421 ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg catcatatac gcagcgaccg
      481 catgacgcaa gctgttttac tcaaatacac atcacctttt tagacggcgg cgctcggttt
      541 cttcagcggc caagctggcc ggccaggccg ccagcttggc atcagacaaa ccggccagga
      601 tttcatgcag ccgcacggct gactcccgag gggaaccctg tggttggcat gcacatacaa
      661 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc
      721 tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaacggat
      781 cacttcgtgt ccctcggtca catgtgcatc ctctagtagc ggtcctgctg agcctcgaca
      841 tgttgtcgca aaattcgccc tggacccgcc caacgatttg tcgtcactgt caaggtttga
      901 cctgcacttc atttggggcc cacatacacc aaaaaaatgc tgcataattc tcggggcagc
      961 aagtcggtta cccggccgcc gtgctggacc gggttgaatg gtgcccgtaa ctttcggtag
     1021 agcggacggc caatactcaa cttcaaggaa tctcacccat gcgcgccggc ggggaaccgg
     1081 agttcccttc agtgagcgtt attagttcgc cgctcggtgt gtcgtagata ctagcccctg
     1141 gggcactttt gaaatttgaa taagatttat gtaatcagtc ttttaggttt gaccggttct
     1201 gccgcttttt ttaaaattgg atttgtaata ataaaacgca attgtttgtt attgtggcgc
     1261 tctatcatag atgtcgctat aaacctattc agcacaatat attgttttca ttttaatatt
     1321 gtacatataa gtagtagggt acaatcagta aattgaacgg agaatattat tcataaaaat
     1381 acgatagtaa cgggtgatat attcattaga atgaaccgaa accggcggta aggatctgag
     1441 ctacacatgc tcaggttttt tacaacgtgc acaacagaat tgaaagcaaa tatcatgcga
     1501 tcataggctt ctcgcatatc tcattaaagc aggacaagct cagatccact cctcagggat
     1561 cttcttaggc ttctttccgt ccaagaagtt agccttccac ttctggaaga aagggttgtt
     1621 cctgtgagca gtgtcgtgag ctccgtccat tcccaccttc ttagcgaact tctccaaaga
     1681 caagttctga gccatcaaca ctccagcgat cttctcctgc acgttgtcag aaagcttagc
     1741 gatctccaca gggatagcct cgtgagactt caccaacttc cacaacttgt cagagttcaa
     1801 tcctcctccc attgtatcga taattgtaaa tgtaattgta atgttgtttg ttgtttgttg
     1861 ttgttggtaa ttgttgtaaa aatgagctct tatactcgag cgtgtcctct ccaaatgaaa
     1921 tgaacttcct tatatagagg aagggtcttg cgaaggatag tgggattgtg cgtcatccct
     1981 tacgtcagtg gagatgtcac atcaatccac ttgctttgta gacgtggttg gaacctcttc
     2041 tttttccacg atgctcctcg tgggtggggg tccatctttg ggaccactgt cggcagagag
     2101 atcttgaatg atagcctttc ctttatcgca atgatggcat ttgtaggagc caccttcctt
     2161 ttctactgtc ctttcgatga agtgacagat agctgggcaa tggaatccga ggaggtttcc
     2221 cgaaattatc ctttgttgaa aagtctcaat agccctttga tcttctgaga ctgtatcttt
     2281 gacatttttg gagtagacca gagtgtcgtg ctccaccatg ttgacgaaga ttttcttctt
     2341 gtcattgagt cgtaaaagac tctgtatgaa ctgttcgcca gtcttgacgg cgagttctgt
     2401 tagatcctcg atttgaatct tagactccat gcatggcctt agattcagta ggaactacct
     2461 ttttagagac tccaatctct attacttgcc ttggtttatg aagcaagcct tgaatcgtcc
     2521 atactggaat agtacttctg atcttgagaa atatgtcttt ctctgtgttc ttgatgcaat
     2581 tagtcctgaa tcttttgact gcatctttaa ccttcttggg aaggtatttg atctcctgga
     2641 gattgttact cgggtagatc gtcttgatga gacttgctgc gtaggcctct ctaaccatct
     2701 gtgggtcagc attctttctg aaattgaaga ggctaacctt ctcattatca gtggtgaaca
     2761 tagtgtcgtc accttcacct tcgaacttcc ttcctagatc gtaaagatag aggaaatcgt
     2821 ccattgtaat ctccggggca aaggagatct cttttggggc tggatcactg ctgggccttt
     2881 tggttcctag cgtgagccag tgggcttttt gctttggtgg gcttgttagg gccttagcaa
     2941 agctcttggg cttgagttga gcttctcctt tggggatgaa gttcaacctg tctgtttgct
     3001 gacttgttgt gtacgcgtca gctgctgctc ttgcctctgt aatagtggca aatttcttgt
     3061 gtgcaactcc gggaacgccg tttgttgccg cctttgtaca accccagtca tcgtatatac
     3121 cggcatgtgg accgttatac acaacgtagt agttgatatg agggtgttga atacccgatt
     3181 ctgctctgag aggagcaact gtgctgttaa gctcagattt ttgtgggatt ggaattcctc
     3241 caagcttgaa ttcttgcagc gctccaattc agcaagcgtc gatctagtaa catagatgac
     3301 accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat cgcgtattaa
     3361 atgtataatt gcgggactct aatcataaaa acccatctca taaataacgt catgcattac
     3421 atgttaatta ttacatgctt aacgtaattc aacagaaatt atatgataat catcgcaaga
     3481 ccggcaacag gattcaatct taagaaactt tattgccaaa tgtttgaacg atctgcttga
     3541 ctctagatcc agagtcccgc tcaaatctca gtcacaggaa gaacaggtct aggaggaaca
     3601 ggaagagaga aatcaagctg ccagaatccc acatcatgcc aatttccatg tttgaaacca
     3661 gcagctctaa gcattcccct aggagcataa ccaagagcct catgcatcct cacagaagga
     3721 tcattaggaa gtccaatcac agcaaccaca gacttgaaac cctgagcctc caaagacttc
     3781 aaaagatgag tgtaaagagt agatccaaga ccagttcttt gatgcctagg agacacgtaa
     3841 acagtagact cagcagtcca atcgtaagca ttcctagcct tccaaggacc agcataagca
     3901 ataccagcca cttcaccatc cacttcagca acaagccaag gatacctctc tctaagcctc
     3961 acaagatcat cagtccactc ttgtggttct tgaggctcag tcctgaaatt cacagtagaa
     4021 gtctcaatgt aatgattcac aatagtgcac acagcaggca tatcagcctc agtagccctt
     4081 ctaatatcag caggccttct ttcaggagac atgcgaaacg atccagatcc ggtgcagatt
     4141 atttggattg agagtgaata tgagactcta attggatacc gaggggaatt tatggaacgt
     4201 cagtggagca ttttgacaag aaatatttgc tagctgatag tgaccttagg cgacttttga
     4261 acgcgcaata atggtttctg acgtatgtgc ttagctcatt aaactccaga aacccgcggc
     4321 tgagtggctc cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg cttgtcccgc
     4381 gtcatcggcg ggggtcataa cgtgactccc ttaattctcc gctccggatc cgaattcggc
     4441 acagagtgtt caaccccagt acattaaaaa cgtccgcaat gtgttattaa gttgtctaag
     4501 cgtcaatttg tttacaccac aatatatcct gccaccagcc agccaacagc tccccgaccg
     4561 gcagctcggc acaaaatcac cactcgatac aggcagccca tcaaccatga ctactgtccg
     4621 gtccaacaga tccgagcagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5101 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5341 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5401 gtatatatga gtaaacttgg tctgacagct aaaacaattc atccagtaaa atataatatt
     5461 ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg acatactgtt
     5521 cttccccgat atcctccctg agcgaccgga cgcagaaggc aatgtcatac cacttgtccg
     5581 ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc acaaagatgt
     5641 tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt tccgtcttta
     5701 aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ctcttcccag ttttcgcaat
     5761 ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg ctgtctaagc
     5821 tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg cactccgcat
     5881 acagctcgat aatcttttca gggctttgtt catcttcata ctcttccgag caaaggacgc
     5941 catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag tgcaggacct
     6001 ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt tccacatcat
     6061 aggtggtccc tttataccgg ctgtccgtca tttttaaata taggttttca ttttctccca
     6121 ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag cggtattttt
     6181 cgatcagttt tttcaattcc ggtgatattc tcattttagc catactcttc ctttttcaat
     6241 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     6301 agaaaaataa acaaataggg gttccgcgca ctcggagcct tgatgtgggc gctttttgca
     6361 gctgttcggc tgtgcgctgg ccagacagtt atgcacaggc caggcgggtt ttaagagttt
     6421 taataagttt taaagagttt taggcggaaa aatcgccttt tttctctttt atatcagtca
     6481 cttacatgtg tgaccggttc ccaatgtacg gctttgggtt cccaatgtac gggttccggt
     6541 tcccaatgta cggctttggg ttcccaatgt acgtgctatc cacaggaaag aagctttttc
     6601 gacctttttc ccctgctagg gcaatttgcc ctagcatctg ctccgtacat taggaaccgg
     6661 cggatgcttc gccctcgatc aggttgcggt agcgcatgac taggatcggg ccagcctgcc
     6721 ccgcctcctc cttcaaatcg tactccggca ggtcatttga cccgatcagc ttgcgcacgg
     6781 tgaaacagaa cttcttgaac tctccggcgc tgccactgcg ttcgtagatc gtcttgaaca
     6841 accatctggc ttctgccttg cctgcggcgc ggcgtgccag gcggtagaga aaacggccga
     6901 tgccgggatc gatcaaaaag taatcggggt gaaccgtcag cacgtccggg ttcttgcctt
     6961 ctgtgatctc gcggtacatc caatcagcta gctcgatctc gatgtactcc ggccgcccgg
     7021 tttcgctctt tacgatcttg tagcggctaa tcaaggcttc accctcggat accgtcacca
     7081 ggcggccgtt cttggccttc ttcgtacgct gcatggcaac gtgcgtggtg tttaaccgaa
     7141 tgcaggtttc taccaggtcg tctttctgct ttccgccatc ggctcgccgg cagaacttga
     7201 gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt gtctcccttc ccttcccggt
     7261 atcggttcat ggattcggtt agatgggaaa ccgccatcag taccaggtcg taatcccaca
     7321 cactggccat gccggccggc cctgcggaaa cctctacgtg cccgtctgga agctcgtagc
     7381 ggatcacctc gccagctcgt cggtcacgct tcgacagacg gaaaacggcc acgtccatga
     7441 tgctgcgact atcgcgggtg cccacgtcat agagcatcgg aacgaaaaaa tctggttgct
     7501 cgtcgccctt gggcggcttc ctaatcgacg acgcaccggc tgccggcggt tgccgggatt
     7561 ctttgcggat tcgatcagcg gccgcttgcc acgattcacc ggggcgtgct tctgcctcga
     7621 tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc caccaggtca tcacccagcg
     7681 ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct cacgccgatt cctcgggctt
     7741 gggggttcca gtgccattgc agggccggca gacaacccag ccgcttacgc ctggccaacc
     7801 gcccgttcct ccacacatgg ggcattccac ggcgtcggtg cctggttgtt cttgattttc
     7861 catgccgcct cctttagccg ctaaaattca tctactcatt tattcatttg ctcatttact
     7921 ctggtagctg cgcgatgtat tcagatagca gctcggtaat ggtcttgcct tggcgtaccg
     7981 cgtacatctt cagcttggtg tgatcctccg ccggcaactg aaagttgacc cgcttcatgg
     8041 ctggcgtgtc tgccaggctg gccaacgttg cagccttgct gctgcgtgcg ctcggacggc
     8101 cggcacttag cgtgtttgtg cttttgctca ttttctcttt acctcattaa ctcaaatgag
     8161 ttttgattta atttcagcgg ccagcgcctg gacctcgcgg gcagcgtcgc cctcgggttc
     8221 tgattcaaga acggttgtgc cggcggcggc agtgcctggg tagctcacgc gctgcgtgat
     8281 acgggactca agaatgggca gctcgtaccc ggccagcgcc tcggcaacct caccgccgat
     8341 gcgcgtgcct ttgatc
//
