LOCUS       sequence-342139-        8398 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 8398)
  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 8398)
  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_04266a
FEATURES             Location/Qualifiers
     source          1..8398
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    704..728
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(795..1502)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1915..2260)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1952..1972)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3293..3545)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4145..4327)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4181..4359)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4522..4546
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4682..5270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4762..4781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5441..6235)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6236..6340)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6402..6596
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6662..7735)
                     /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(8164..8398,1..395))
                     /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 cttccatccg tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat
       61 atgtcgtaag ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac
      121 acagccaagt ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg
      181 gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct
      241 tcccgcacgg ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca
      301 tcggccccgg cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac
      361 ccgcctttct ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat
      421 ttactcatcg catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac
      481 atcacctttt tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg
      541 ccagcttggc atcagacaaa ccggccagga tttcatgcag ccgcacggct gactcccgag
      601 gggaaccctg tggttggcat gcacatacaa atggacgaac ggataaacct tttcacgccc
      661 ttttaaatat ccgattattc taataaacgc tcttttctct taggtttacc cgccaatata
      721 tcctgtcaaa cactgatagt ttaaacggat cacttcgtgt ccctcggtca catgtgcatc
      781 ctctagtagc ggtcctgctg agcctcgaca tgttgtcgca aaattcgccc tggacccgcc
      841 caacgatttg tcgtcactgt caaggtttga cctgcacttc atttggggcc cacatacacc
      901 aaaaaaatgc tgcataattc tcggggcagc aagtcggtta cccggccgcc gtgctggacc
      961 gggttgaatg gtgcccgtaa ctttcggtag agcggacggc caatactcaa cttcaaggaa
     1021 tctcacccat gcgcgccggc ggggaaccgg agttcccttc agtgagcgtt attagttcgc
     1081 cgctcggtgt gtcgtagata ctagcccctg gggcactttt gaaatttgaa taagatttat
     1141 gtaatcagtc ttttaggttt gaccggttct gccgcttttt ttaaaattgg atttgtaata
     1201 ataaaacgca attgtttgtt attgtggcgc tctatcatag atgtcgctat aaacctattc
     1261 agcacaatat attgttttca ttttaatatt gtacatataa gtagtagggt acaatcagta
     1321 aattgaacgg agaatattat tcataaaaat acgatagtaa cgggtgatat attcattaga
     1381 atgaaccgaa accggcggta aggatctgag ctacacatgc tcaggttttt tacaacgtgc
     1441 acaacagaat tgaaagcaaa tatcatgcga tcataggctt ctcgcatatc tcattaaagc
     1501 aggacaagct cagtcagaga tcctgtcgat ccatccaagc aatcctcccc tagtaggctt
     1561 tggtgtaggt gttggagtag gagttgcagt agagttcaaa gcaggtgttg gagttggtgt
     1621 ctccttgtgc ttcttctttt tcttctttct cttcctcctc ttgaacaatc ccctctcgtc
     1681 gttgtcgaaa gaagagttct cgtcctcgtc ctcgtactcg ttatcctcgt ccttccacaa
     1741 catgtcgtcg tcgtcgtgca aagtgtcgaa cacgtatcca gtaggagcct tagtagcctc
     1801 gtaagtctta gactcagaga tcattgtatc gataattgta aatgtaattg taatgttgtt
     1861 tgttgtttgt tgttgttggt aattgttgta aaaatgagct cttatactcg agcgtgtcct
     1921 ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg
     1981 tgcgtcatcc cttacgtcag tggagatgtc acatcaatcc acttgctttg tagacgtggt
     2041 tggaacctct tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact
     2101 gtcggcagag agatcttgaa tgatagcctt tcctttatcg caatgatggc atttgtagga
     2161 gccaccttcc ttttctactg tcctttcgat gaagtgacag atagctgggc aatggaatcc
     2221 gaggaggttt cccgaaatta tcctttgttg aaaagtctca atagcccttt gatcttctga
     2281 gactgtatct ttgacatttt tggagtagac cagagtgtcg tgctccacca tgttgacgaa
     2341 gattttcttc ttgtcattga gtcgtaaaag actctgtatg aactgttcgc cagtcttgac
     2401 ggcgagttct gttagatcct cgatttgaat cttagactcc atgcatggcc ttagattcag
     2461 taggaactac ctttttagag actccaatct ctattacttg ccttggttta tgaagcaagc
     2521 cttgaatcgt ccatactgga atagtacttc tgatcttgag aaatatgtct ttctctgtgt
     2581 tcttgatgca attagtcctg aatcttttga ctgcatcttt aaccttcttg ggaaggtatt
     2641 tgatctcctg gagattgtta ctcgggtaga tcgtcttgat gagacttgct gcgtaggcct
     2701 ctctaaccat ctgtgggtca gcattctttc tgaaattgaa gaggctaacc ttctcattat
     2761 cagtggtgaa catagtgtcg tcaccttcac cttcgaactt ccttcctaga tcgtaaagat
     2821 agaggaaatc gtccattgta atctccgggg caaaggagat ctcttttggg gctggatcac
     2881 tgctgggcct tttggttcct agcgtgagcc agtgggcttt ttgctttggt gggcttgtta
     2941 gggccttagc aaagctcttg ggcttgagtt gagcttctcc tttggggatg aagttcaacc
     3001 tgtctgtttg ctgacttgtt gtgtacgcgt cagctgctgc tcttgcctct gtaatagtgg
     3061 caaatttctt gtgtgcaact ccgggaacgc cgtttgttgc cgcctttgta caaccccagt
     3121 catcgtatat accggcatgt ggaccgttat acacaacgta gtagttgata tgagggtgtt
     3181 gaatacccga ttctgctctg agaggagcaa ctgtgctgtt aagctcagat ttttgtggga
     3241 ttggaattcc tccaagcttg aattcttgca gcgctccaat tcagcaagcg tcgatctagt
     3301 aacatagatg acaccgcgcg cgataattta tcctagtttg cgcgctatat tttgttttct
     3361 atcgcgtatt aaatgtataa ttgcgggact ctaatcataa aaacccatct cataaataac
     3421 gtcatgcatt acatgttaat tattacatgc ttaacgtaat tcaacagaaa ttatatgata
     3481 atcatcgcaa gaccggcaac aggattcaat cttaagaaac tttattgcca aatgtttgaa
     3541 cgatctgctt gactctagat ccagagtccc gctcaaatct cagtcacagg aagaacaggt
     3601 ctaggaggaa caggaagaga gaaatcaagc tgccagaatc ccacatcatg ccaatttcca
     3661 tgtttgaaac cagcagctct aagcattccc ctaggagcat aaccaagagc ctcatgcatc
     3721 ctcacagaag gatcattagg aagtccaatc acagcaacca cagacttgaa accctgagcc
     3781 tccaaagact tcaaaagatg agtgtaaaga gtagatccaa gaccagttct ttgatgccta
     3841 ggagacacgt aaacagtaga ctcagcagtc caatcgtaag cattcctagc cttccaagga
     3901 ccagcataag caataccagc cacttcacca tccacttcag caacaagcca aggatacctc
     3961 tctctaagcc tcacaagatc atcagtccac tcttgtggtt cttgaggctc agtcctgaaa
     4021 ttcacagtag aagtctcaat gtaatgattc acaatagtgc acacagcagg catatcagcc
     4081 tcagtagccc ttctaatatc agcaggcctt ctttcaggag acatgcgaaa cgatccagat
     4141 ccggtgcaga ttatttggat tgagagtgaa tatgagactc taattggata ccgaggggaa
     4201 tttatggaac gtcagtggag cattttgaca agaaatattt gctagctgat agtgacctta
     4261 ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt gcttagctca ttaaactcca
     4321 gaaacccgcg gctgagtggc tccttcaacg ttgcggttct gtcagttcca aacgtaaaac
     4381 ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc ccttaattct ccgctccgga
     4441 tccgaattcg gcacagagtg ttcaacccca gtacattaaa aacgtccgca atgtgttatt
     4501 aagttgtcta agcgtcaatt tgtttacacc acaatatatc ctgccaccag ccagccaaca
     4561 gctccccgac cggcagctcg gcacaaaatc accactcgat acaggcagcc catcaaccat
     4621 gactactgtc cggtccaaca gatccgagca ggaaccgtaa aaaggccgcg ttgctggcgt
     4681 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4741 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4801 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4861 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4921 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4981 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5041 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5101 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5161 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5221 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5281 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5341 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5401 aatcaatcta aagtatatat gagtaaactt ggtctgacag ctaaaacaat tcatccagta
     5461 aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca aaaaatagct
     5521 cgacatactg ttcttccccg atatcctccc tgagcgaccg gacgcagaag gcaatgtcat
     5581 accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact ttgccatctt
     5641 tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc tcttcgggct
     5701 tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg tcctcttccc
     5761 agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat tcggctaagc
     5821 ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga aagagcctga
     5881 tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca tactcttccg
     5941 agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca tgccgttcaa
     6001 agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg tccttttccc
     6061 gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa tataggtttt
     6121 cattttctcc caccagctta tataccttag caggagacat tccttccgta tcttttacgc
     6181 agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta gccatactct
     6241 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6301 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cactcggagc cttgatgtgg
     6361 gcgctttttg cagctgttcg gctgtgcgct ggccagacag ttatgcacag gccaggcggg
     6421 ttttaagagt tttaataagt tttaaagagt tttaggcgga aaaatcgcct tttttctctt
     6481 ttatatcagt cacttacatg tgtgaccggt tcccaatgta cggctttggg ttcccaatgt
     6541 acgggttccg gttcccaatg tacggctttg ggttcccaat gtacgtgcta tccacaggaa
     6601 agaagctttt tcgacctttt tcccctgcta gggcaatttg ccctagcatc tgctccgtac
     6661 attaggaacc ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg actaggatcg
     6721 ggccagcctg ccccgcctcc tccttcaaat cgtactccgg caggtcattt gacccgatca
     6781 gcttgcgcac ggtgaaacag aacttcttga actctccggc gctgccactg cgttcgtaga
     6841 tcgtcttgaa caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga
     6901 gaaaacggcc gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg
     6961 ggttcttgcc ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc tcgatgtact
     7021 ccggccgccc ggtttcgctc tttacgatct tgtagcggct aatcaaggct tcaccctcgg
     7081 ataccgtcac caggcggccg ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg
     7141 tgtttaaccg aatgcaggtt tctaccaggt cgtctttctg ctttccgcca tcggctcgcc
     7201 ggcagaactt gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc ttgtctccct
     7261 tcccttcccg gtatcggttc atggattcgg ttagatggga aaccgccatc agtaccaggt
     7321 cgtaatccca cacactggcc atgccggccg gccctgcgga aacctctacg tgcccgtctg
     7381 gaagctcgta gcggatcacc tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg
     7441 ccacgtccat gatgctgcga ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa
     7501 aatctggttg ctcgtcgccc ttgggcggct tcctaatcga cgacgcaccg gctgccggcg
     7561 gttgccggga ttctttgcgg attcgatcag cggccgcttg ccacgattca ccggggcgtg
     7621 cttctgcctc gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt
     7681 catcacccag cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg ctcacgccga
     7741 ttcctcgggc ttgggggttc cagtgccatt gcagggccgg cagacaaccc agccgcttac
     7801 gcctggccaa ccgcccgttc ctccacacat ggggcattcc acggcgtcgg tgcctggttg
     7861 ttcttgattt tccatgccgc ctcctttagc cgctaaaatt catctactca tttattcatt
     7921 tgctcattta ctctggtagc tgcgcgatgt attcagatag cagctcggta atggtcttgc
     7981 cttggcgtac cgcgtacatc ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga
     8041 cccgcttcat ggctggcgtg tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg
     8101 cgctcggacg gccggcactt agcgtgtttg tgcttttgct cattttctct ttacctcatt
     8161 aactcaaatg agttttgatt taatttcagc ggccagcgcc tggacctcgc gggcagcgtc
     8221 gccctcgggt tctgattcaa gaacggttgt gccggcggcg gcagtgcctg ggtagctcac
     8281 gcgctgcgtg atacgggact caagaatggg cagctcgtac ccggccagcg cctcggcaac
     8341 ctcaccgccg atgcgcgtgc ctttgatcgc ccgcgacacg acaaaggccg cttgtagc
//
