LOCUS       sequence-342347-        9484 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 9484)
  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 9484)
  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_15123
FEATURES             Location/Qualifiers
     source          1..9484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    632..656
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(723..1430)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2929..3274)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2966..2986)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4307..4559)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5159..5341)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5195..5373)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5536..5560
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5696..6284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5776..5795)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6455..7249)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7250..7354)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7416..7610
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7676..8749)
                     /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(9178..9484,1..323))
                     /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 cttggctgca ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc
       61 gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg
      121 cggtcaatcg tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc
      181 gcccaatcgc gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg
      241 agttgcaggg cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg
      301 ttaagtacag cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca
      361 tcatatacgc agcgaccgca tgacgcaagc tgttttactc aaatacacat caccttttta
      421 gacggcggcg ctcggtttct tcagcggcca agctggccgg ccaggccgcc agcttggcat
      481 cagacaaacc ggccaggatt tcatgcagcc gcacggctga ctcccgaggg gaaccctgtg
      541 gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc
      601 gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca
      661 ctgatagttt aaacggatca cttcgtgtcc ctcggtcaca tgtgcatcct ctagtagcgg
      721 tcctgctgag cctcgacatg ttgtcgcaaa attcgccctg gacccgccca acgatttgtc
      781 gtcactgtca aggtttgacc tgcacttcat ttggggccca catacaccaa aaaaatgctg
      841 cataattctc ggggcagcaa gtcggttacc cggccgccgt gctggaccgg gttgaatggt
      901 gcccgtaact ttcggtagag cggacggcca atactcaact tcaaggaatc tcacccatgc
      961 gcgccggcgg ggaaccggag ttcccttcag tgagcgttat tagttcgccg ctcggtgtgt
     1021 cgtagatact agcccctggg gcacttttga aatttgaata agatttatgt aatcagtctt
     1081 ttaggtttga ccggttctgc cgcttttttt aaaattggat ttgtaataat aaaacgcaat
     1141 tgtttgttat tgtggcgctc tatcatagat gtcgctataa acctattcag cacaatatat
     1201 tgttttcatt ttaatattgt acatataagt agtagggtac aatcagtaaa ttgaacggag
     1261 aatattattc ataaaaatac gatagtaacg ggtgatatat tcattagaat gaaccgaaac
     1321 cggcggtaag gatctgagct acacatgctc aggtttttta caacgtgcac aacagaattg
     1381 aaagcaaata tcatgcgatc ataggcttct cgcatatctc attaaagcag gacaagctca
     1441 cctccttggg ttgattgatg tgtcgatgtg aggaggtgcc attttcttct tgtaatagtc
     1501 cttgtatcta gcagcagtag ccctctccaa gttagatgct gtgttctctg atgatctgaa
     1561 aattgtacca taaatatcag aaggctctgt ctccctcttg aaccacctac cgaacaacat
     1621 cctttggtac tcagtagcga ttgagatttc gtccaagtca gcgtgtccgt tcctaatcaa
     1681 gtgctccctt gccctagttg tctgaattgc caactcgtcc tcacctccga aacccttaga
     1741 cactgccctc aacataccct cgttgtacaa aaggttcttg ttgttcaaag tagtcaaata
     1801 accagaaaga gtgtcgaact ttggattagt aggagaaagc aaatcgttaa agtgaatgtc
     1861 caaaagtctg aacacagagt ttgcagagat tccaccctct ttccacttca agaattgttg
     1921 cttttgcaat gtgataacca agttcattgt agctgcgtcg tcaggcttag cgaatgtaag
     1981 cacagttgcc actgctttat ttccgtactg agccacaaat gagtcgatca aagatgcctt
     2041 tgaatttggg ttttttgcgt tatacattga gatgaaagct ccgaaagtgt gcacgttcct
     2101 gtcaagcaat gcgtccaatc ccctgttcaa cctcaacttt ttgtagatat gtttaggtct
     2161 aagaagctcc accctcaccc acttgttgat aagagcgttc tgcaaagctg aagcaagctc
     2221 ccttgtctta acgtcctccc ttgctctagc aaggattgtt gcaagctgct cctctccacc
     2281 aaatccttta gtcaaagttt tcaaaagtga gtctttagta gactttgttg aaataaccaa
     2341 tttaatgtag tcgtcaagca cttccatctt aggagaagcc aatgccaaat atccatcaga
     2401 cctcaattta agcaaagtaa aaacgtcatc aactgatttc ctcttctgca accaagaaag
     2461 caactgagcg tgcctaagcc tctggatagt gttcatctca ccaccagccc tctgtgcagt
     2521 catcaatgcg tttgccacag cgtcgtcacc gtagtgagcc atgaacatac cgatcactga
     2581 agccttgttg ttgttgttct gagtattcaa ctctttaaca taattttcca atgccttaag
     2641 gttaggtgaa gcaagagcgt cgtccattcc gtttgtcaac ttgaacctcc tcaagatagg
     2701 ctcagctgga gggtccttag agatccatga tccaaggttg atcttagaca ctccagcctt
     2761 agcaaggtct gtaagcttct ccaatcctgc tgagatcacc ctgttctcac cattctcagt
     2821 gttctgtgta gacaccattg tatcgataat tgtaaatgta attgtaatgt tgtttgttgt
     2881 ttgttgttgt tggtaattgt tgtaaaaatg agctcttata ctcgagcgtg tcctctccaa
     2941 atgaaatgaa cttccttata tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc
     3001 atcccttacg tcagtggaga tgtcacatca atccacttgc tttgtagacg tggttggaac
     3061 ctcttctttt tccacgatgc tcctcgtggg tgggggtcca tctttgggac cactgtcggc
     3121 agagagatct tgaatgatag cctttccttt atcgcaatga tggcatttgt aggagccacc
     3181 ttccttttct actgtccttt cgatgaagtg acagatagct gggcaatgga atccgaggag
     3241 gtttcccgaa attatccttt gttgaaaagt ctcaatagcc ctttgatctt ctgagactgt
     3301 atctttgaca tttttggagt agaccagagt gtcgtgctcc accatgttga cgaagatttt
     3361 cttcttgtca ttgagtcgta aaagactctg tatgaactgt tcgccagtct tgacggcgag
     3421 ttctgttaga tcctcgattt gaatcttaga ctccatgcat ggccttagat tcagtaggaa
     3481 ctaccttttt agagactcca atctctatta cttgccttgg tttatgaagc aagccttgaa
     3541 tcgtccatac tggaatagta cttctgatct tgagaaatat gtctttctct gtgttcttga
     3601 tgcaattagt cctgaatctt ttgactgcat ctttaacctt cttgggaagg tatttgatct
     3661 cctggagatt gttactcggg tagatcgtct tgatgagact tgctgcgtag gcctctctaa
     3721 ccatctgtgg gtcagcattc tttctgaaat tgaagaggct aaccttctca ttatcagtgg
     3781 tgaacatagt gtcgtcacct tcaccttcga acttccttcc tagatcgtaa agatagagga
     3841 aatcgtccat tgtaatctcc ggggcaaagg agatctcttt tggggctgga tcactgctgg
     3901 gccttttggt tcctagcgtg agccagtggg ctttttgctt tggtgggctt gttagggcct
     3961 tagcaaagct cttgggcttg agttgagctt ctcctttggg gatgaagttc aacctgtctg
     4021 tttgctgact tgttgtgtac gcgtcagctg ctgctcttgc ctctgtaata gtggcaaatt
     4081 tcttgtgtgc aactccggga acgccgtttg ttgccgcctt tgtacaaccc cagtcatcgt
     4141 atataccggc atgtggaccg ttatacacaa cgtagtagtt gatatgaggg tgttgaatac
     4201 ccgattctgc tctgagagga gcaactgtgc tgttaagctc agatttttgt gggattggaa
     4261 ttcctccaag cttgaattct tgcagcgctc caattcagca agcgtcgatc tagtaacata
     4321 gatgacaccg cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg
     4381 tattaaatgt ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg
     4441 cattacatgt taattattac atgcttaacg taattcaaca gaaattatat gataatcatc
     4501 gcaagaccgg caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct
     4561 gcttgactct agatccagag tcccgctcaa atctcagtca caggaagaac aggtctagga
     4621 ggaacaggaa gagagaaatc aagctgccag aatcccacat catgccaatt tccatgtttg
     4681 aaaccagcag ctctaagcat tcccctagga gcataaccaa gagcctcatg catcctcaca
     4741 gaaggatcat taggaagtcc aatcacagca accacagact tgaaaccctg agcctccaaa
     4801 gacttcaaaa gatgagtgta aagagtagat ccaagaccag ttctttgatg cctaggagac
     4861 acgtaaacag tagactcagc agtccaatcg taagcattcc tagccttcca aggaccagca
     4921 taagcaatac cagccacttc accatccact tcagcaacaa gccaaggata cctctctcta
     4981 agcctcacaa gatcatcagt ccactcttgt ggttcttgag gctcagtcct gaaattcaca
     5041 gtagaagtct caatgtaatg attcacaata gtgcacacag caggcatatc agcctcagta
     5101 gcccttctaa tatcagcagg ccttctttca ggagacatgc gaaacgatcc agatccggtg
     5161 cagattattt ggattgagag tgaatatgag actctaattg gataccgagg ggaatttatg
     5221 gaacgtcagt ggagcatttt gacaagaaat atttgctagc tgatagtgac cttaggcgac
     5281 ttttgaacgc gcaataatgg tttctgacgt atgtgcttag ctcattaaac tccagaaacc
     5341 cgcggctgag tggctccttc aacgttgcgg ttctgtcagt tccaaacgta aaacggcttg
     5401 tcccgcgtca tcggcggggg tcataacgtg actcccttaa ttctccgctc cggatccgaa
     5461 ttcggcacag agtgttcaac cccagtacat taaaaacgtc cgcaatgtgt tattaagttg
     5521 tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc
     5581 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcaa ccatgactac
     5641 tgtccggtcc aacagatccg agcaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5701 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5761 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5821 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5881 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5941 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     6001 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     6061 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     6121 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     6181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     6241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     6301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     6361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     6421 tctaaagtat atatgagtaa acttggtctg acagctaaaa caattcatcc agtaaaatat
     6481 aatattttat tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat agctcgacat
     6541 actgttcttc cccgatatcc tccctgagcg accggacgca gaaggcaatg tcataccact
     6601 tgtccgccct gccgcttctc ccaagatcaa taaagccact tactttgcca tctttcacaa
     6661 agatgttgct gtctcccagg tcgccgtggg aaaagacaag ttcctcttcg ggcttttccg
     6721 tctttaaaaa atcatacagc tcgcgcggat ctttaaatgg agtgtcctct tcccagtttt
     6781 cgcaatccac atcggccaga tcgttattca gtaagtaatc caattcggct aagcggctgt
     6841 ctaagctatt cgtataggga caatccgata tgtcgatgga gtgaaagagc ctgatgcact
     6901 ccgcatacag ctcgataatc ttttcagggc tttgttcatc ttcatactct tccgagcaaa
     6961 ggacgccatc ggcctcactc atgagcagat tgctccagcc atcatgccgt tcaaagtgca
     7021 ggacctttgg aacaggcagc tttccttcca gccatagcat catgtccttt tcccgttcca
     7081 catcataggt ggtcccttta taccggctgt ccgtcatttt taaatatagg ttttcatttt
     7141 ctcccaccag cttatatacc ttagcaggag acattccttc cgtatctttt acgcagcggt
     7201 atttttcgat cagttttttc aattccggtg atattctcat tttagccata ctcttccttt
     7261 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7321 gtatttagaa aaataaacaa ataggggttc cgcgcactcg gagccttgat gtgggcgctt
     7381 tttgcagctg ttcggctgtg cgctggccag acagttatgc acaggccagg cgggttttaa
     7441 gagttttaat aagttttaaa gagttttagg cggaaaaatc gccttttttc tcttttatat
     7501 cagtcactta catgtgtgac cggttcccaa tgtacggctt tgggttccca atgtacgggt
     7561 tccggttccc aatgtacggc tttgggttcc caatgtacgt gctatccaca ggaaagaagc
     7621 tttttcgacc tttttcccct gctagggcaa tttgccctag catctgctcc gtacattagg
     7681 aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg catgactagg atcgggccag
     7741 cctgccccgc ctcctccttc aaatcgtact ccggcaggtc atttgacccg atcagcttgc
     7801 gcacggtgaa acagaacttc ttgaactctc cggcgctgcc actgcgttcg tagatcgtct
     7861 tgaacaacca tctggcttct gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac
     7921 ggccgatgcc gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg tccgggttct
     7981 tgccttctgt gatctcgcgg tacatccaat cagctagctc gatctcgatg tactccggcc
     8041 gcccggtttc gctctttacg atcttgtagc ggctaatcaa ggcttcaccc tcggataccg
     8101 tcaccaggcg gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta
     8161 accgaatgca ggtttctacc aggtcgtctt tctgctttcc gccatcggct cgccggcaga
     8221 acttgagtac gtccgcaacg tgtggacgga acacgcggcc gggcttgtct cccttccctt
     8281 cccggtatcg gttcatggat tcggttagat gggaaaccgc catcagtacc aggtcgtaat
     8341 cccacacact ggccatgccg gccggccctg cggaaacctc tacgtgcccg tctggaagct
     8401 cgtagcggat cacctcgcca gctcgtcggt cacgcttcga cagacggaaa acggccacgt
     8461 ccatgatgct gcgactatcg cgggtgccca cgtcatagag catcggaacg aaaaaatctg
     8521 gttgctcgtc gcccttgggc ggcttcctaa tcgacgacgc accggctgcc ggcggttgcc
     8581 gggattcttt gcggattcga tcagcggccg cttgccacga ttcaccgggg cgtgcttctg
     8641 cctcgatgcg ttgccgctgg gcggcctgcg cggccttcaa cttctccacc aggtcatcac
     8701 ccagcgccgc gccgatttgt accgggccgg atggtttgcg accgctcacg ccgattcctc
     8761 gggcttgggg gttccagtgc cattgcaggg ccggcagaca acccagccgc ttacgcctgg
     8821 ccaaccgccc gttcctccac acatggggca ttccacggcg tcggtgcctg gttgttcttg
     8881 attttccatg ccgcctcctt tagccgctaa aattcatcta ctcatttatt catttgctca
     8941 tttactctgg tagctgcgcg atgtattcag atagcagctc ggtaatggtc ttgccttggc
     9001 gtaccgcgta catcttcagc ttggtgtgat cctccgccgg caactgaaag ttgacccgct
     9061 tcatggctgg cgtgtctgcc aggctggcca acgttgcagc cttgctgctg cgtgcgctcg
     9121 gacggccggc acttagcgtg tttgtgcttt tgctcatttt ctctttacct cattaactca
     9181 aatgagtttt gatttaattt cagcggccag cgcctggacc tcgcgggcag cgtcgccctc
     9241 gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg
     9301 cgtgatacgg gactcaagaa tgggcagctc gtacccggcc agcgcctcgg caacctcacc
     9361 gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta gccttccatc
     9421 cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc atatgtcgta
     9481 aggg
//
