LOCUS       sequence-342200-        8593 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 8593)
  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 8593)
  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_07550a
FEATURES             Location/Qualifiers
     source          1..8593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(265..894)
                     /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    1203..1227
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1294..2001)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2609..2954)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2646..2666)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3987..4239)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4839..5021)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4875..5053)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5216..5240
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5376..5964)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5456..5475)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6135..6929)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6930..7034)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7096..7290
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7356..8429)
                     /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 tcatctactc atttattcat ttgctcattt actctggtag ctgcgcgatg tattcagata
       61 gcagctcggt aatggtcttg ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct
      121 ccgccggcaa ctgaaagttg acccgcttca tggctggcgt gtctgccagg ctggccaacg
      181 ttgcagcctt gctgctgcgt gcgctcggac ggccggcact tagcgtgttt gtgcttttgc
      241 tcattttctc tttacctcat taactcaaat gagttttgat ttaatttcag cggccagcgc
      301 ctggacctcg cgggcagcgt cgccctcggg ttctgattca agaacggttg tgccggcggc
      361 ggcagtgcct gggtagctca cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta
      421 cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac
      481 gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa ccagctccac
      541 caggtcggcg gtggcccata tgtcgtaagg gcttggctgc accggaatca gcacgaagtc
      601 ggctgccttg atcgcggaca cagccaagtc cgccgcctgg ggcgctccgt cgatcactac
      661 gaagtcgcgc cggccgatgg ccttcacgtc gcggtcaatc gtcgggcggt cgatgccgac
      721 aacggttagc ggttgatctt cccgcacggc cgcccaatcg cgggcactgc cctggggatc
      781 ggaatcgact aacagaacat cggccccggc gagttgcagg gcgcgggcta gatgggttgc
      841 gatggtcgtc ttgcctgacc cgcctttctg gttaagtaca gcgataacct tcatgcgttc
      901 cccttgcgta tttgtttatt tactcatcgc atcatatacg cagcgaccgc atgacgcaag
      961 ctgttttact caaatacaca tcaccttttt agacggcggc gctcggtttc ttcagcggcc
     1021 aagctggccg gccaggccgc cagcttggca tcagacaaac cggccaggat ttcatgcagc
     1081 cgcacggctg actcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg
     1141 gataaacctt ttcacgccct tttaaatatc cgattattct aataaacgct cttttctctt
     1201 aggtttaccc gccaatatat cctgtcaaac actgatagtt taaacggatc acttcgtgtc
     1261 cctcggtcac atgtgcatcc tctagtagcg gtcctgctga gcctcgacat gttgtcgcaa
     1321 aattcgccct ggacccgccc aacgatttgt cgtcactgtc aaggtttgac ctgcacttca
     1381 tttggggccc acatacacca aaaaaatgct gcataattct cggggcagca agtcggttac
     1441 ccggccgccg tgctggaccg ggttgaatgg tgcccgtaac tttcggtaga gcggacggcc
     1501 aatactcaac ttcaaggaat ctcacccatg cgcgccggcg gggaaccgga gttcccttca
     1561 gtgagcgtta ttagttcgcc gctcggtgtg tcgtagatac tagcccctgg ggcacttttg
     1621 aaatttgaat aagatttatg taatcagtct tttaggtttg accggttctg ccgctttttt
     1681 taaaattgga tttgtaataa taaaacgcaa ttgtttgtta ttgtggcgct ctatcataga
     1741 tgtcgctata aacctattca gcacaatata ttgttttcat tttaatattg tacatataag
     1801 tagtagggta caatcagtaa attgaacgga gaatattatt cataaaaata cgatagtaac
     1861 gggtgatata ttcattagaa tgaaccgaaa ccggcggtaa ggatctgagc tacacatgct
     1921 caggtttttt acaacgtgca caacagaatt gaaagcaaat atcatgcgat cataggcttc
     1981 tcgcatatct cattaaagca ggacaagctc atccagatgg gttcaaagag tgcctctttg
     2041 tcaaatactg agcgttgaac aagtcgtaag cctgctttga cacctcgttc ttagtgatgt
     2101 gcctcaagtc cttaggagtc tttttttgag taatccaaag gttcaccaag aacttcacgt
     2161 cgtcgtctgc gttctcgatc ttcctcaaga tctgagcacc ctccctagtg atcttctggt
     2221 tagcctgagg acctggcttc aaagagtggt gcttcttcat gtactgcacc aagaactctt
     2281 ggtaagcctt cttgttcaca ggagtcttag tcaacaacct caagtcctca ggcctcatgt
     2341 tctgcctcat ccacaagtta gccaagaact tagcgttatc tgcctctccc tgctgcttgt
     2401 tcaatgcagc aagcttcttt gccatagcgt cgtcgttagc aagcaacttc ttagcagcgt
     2461 cgtctgtcct tccaagtccc ttgaatgcgt cgtcgataag agaagtctgg atcttcattg
     2521 tatcgataat tgtaaatgta attgtaatgt tgtttgttgt ttgttgttgt tggtaattgt
     2581 tgtaaaaatg agctcttata ctcgagcgtg tcctctccaa atgaaatgaa cttccttata
     2641 tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc atcccttacg tcagtggaga
     2701 tgtcacatca atccacttgc tttgtagacg tggttggaac ctcttctttt tccacgatgc
     2761 tcctcgtggg tgggggtcca tctttgggac cactgtcggc agagagatct tgaatgatag
     2821 cctttccttt atcgcaatga tggcatttgt aggagccacc ttccttttct actgtccttt
     2881 cgatgaagtg acagatagct gggcaatgga atccgaggag gtttcccgaa attatccttt
     2941 gttgaaaagt ctcaatagcc ctttgatctt ctgagactgt atctttgaca tttttggagt
     3001 agaccagagt gtcgtgctcc accatgttga cgaagatttt cttcttgtca ttgagtcgta
     3061 aaagactctg tatgaactgt tcgccagtct tgacggcgag ttctgttaga tcctcgattt
     3121 gaatcttaga ctccatgcat ggccttagat tcagtaggaa ctaccttttt agagactcca
     3181 atctctatta cttgccttgg tttatgaagc aagccttgaa tcgtccatac tggaatagta
     3241 cttctgatct tgagaaatat gtctttctct gtgttcttga tgcaattagt cctgaatctt
     3301 ttgactgcat ctttaacctt cttgggaagg tatttgatct cctggagatt gttactcggg
     3361 tagatcgtct tgatgagact tgctgcgtag gcctctctaa ccatctgtgg gtcagcattc
     3421 tttctgaaat tgaagaggct aaccttctca ttatcagtgg tgaacatagt gtcgtcacct
     3481 tcaccttcga acttccttcc tagatcgtaa agatagagga aatcgtccat tgtaatctcc
     3541 ggggcaaagg agatctcttt tggggctgga tcactgctgg gccttttggt tcctagcgtg
     3601 agccagtggg ctttttgctt tggtgggctt gttagggcct tagcaaagct cttgggcttg
     3661 agttgagctt ctcctttggg gatgaagttc aacctgtctg tttgctgact tgttgtgtac
     3721 gcgtcagctg ctgctcttgc ctctgtaata gtggcaaatt tcttgtgtgc aactccggga
     3781 acgccgtttg ttgccgcctt tgtacaaccc cagtcatcgt atataccggc atgtggaccg
     3841 ttatacacaa cgtagtagtt gatatgaggg tgttgaatac ccgattctgc tctgagagga
     3901 gcaactgtgc tgttaagctc agatttttgt gggattggaa ttcctccaag cttgaattct
     3961 tgcagcgctc caattcagca agcgtcgatc tagtaacata gatgacaccg cgcgcgataa
     4021 tttatcctag tttgcgcgct atattttgtt ttctatcgcg tattaaatgt ataattgcgg
     4081 gactctaatc ataaaaaccc atctcataaa taacgtcatg cattacatgt taattattac
     4141 atgcttaacg taattcaaca gaaattatat gataatcatc gcaagaccgg caacaggatt
     4201 caatcttaag aaactttatt gccaaatgtt tgaacgatct gcttgactct agatccagag
     4261 tcccgctcaa atctcagtca caggaagaac aggtctagga ggaacaggaa gagagaaatc
     4321 aagctgccag aatcccacat catgccaatt tccatgtttg aaaccagcag ctctaagcat
     4381 tcccctagga gcataaccaa gagcctcatg catcctcaca gaaggatcat taggaagtcc
     4441 aatcacagca accacagact tgaaaccctg agcctccaaa gacttcaaaa gatgagtgta
     4501 aagagtagat ccaagaccag ttctttgatg cctaggagac acgtaaacag tagactcagc
     4561 agtccaatcg taagcattcc tagccttcca aggaccagca taagcaatac cagccacttc
     4621 accatccact tcagcaacaa gccaaggata cctctctcta agcctcacaa gatcatcagt
     4681 ccactcttgt ggttcttgag gctcagtcct gaaattcaca gtagaagtct caatgtaatg
     4741 attcacaata gtgcacacag caggcatatc agcctcagta gcccttctaa tatcagcagg
     4801 ccttctttca ggagacatgc gaaacgatcc agatccggtg cagattattt ggattgagag
     4861 tgaatatgag actctaattg gataccgagg ggaatttatg gaacgtcagt ggagcatttt
     4921 gacaagaaat atttgctagc tgatagtgac cttaggcgac ttttgaacgc gcaataatgg
     4981 tttctgacgt atgtgcttag ctcattaaac tccagaaacc cgcggctgag tggctccttc
     5041 aacgttgcgg ttctgtcagt tccaaacgta aaacggcttg tcccgcgtca tcggcggggg
     5101 tcataacgtg actcccttaa ttctccgctc cggatccgaa ttcggcacag agtgttcaac
     5161 cccagtacat taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc aatttgttta
     5221 caccacaata tatcctgcca ccagccagcc aacagctccc cgaccggcag ctcggcacaa
     5281 aatcaccact cgatacaggc agcccatcaa ccatgactac tgtccggtcc aacagatccg
     5341 agcaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5401 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5461 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5521 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5581 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5641 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5701 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5761 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5821 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5881 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5941 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6001 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6061 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6121 acttggtctg acagctaaaa caattcatcc agtaaaatat aatattttat tttctcccaa
     6181 tcaggcttga tccccagtaa gtcaaaaaat agctcgacat actgttcttc cccgatatcc
     6241 tccctgagcg accggacgca gaaggcaatg tcataccact tgtccgccct gccgcttctc
     6301 ccaagatcaa taaagccact tactttgcca tctttcacaa agatgttgct gtctcccagg
     6361 tcgccgtggg aaaagacaag ttcctcttcg ggcttttccg tctttaaaaa atcatacagc
     6421 tcgcgcggat ctttaaatgg agtgtcctct tcccagtttt cgcaatccac atcggccaga
     6481 tcgttattca gtaagtaatc caattcggct aagcggctgt ctaagctatt cgtataggga
     6541 caatccgata tgtcgatgga gtgaaagagc ctgatgcact ccgcatacag ctcgataatc
     6601 ttttcagggc tttgttcatc ttcatactct tccgagcaaa ggacgccatc ggcctcactc
     6661 atgagcagat tgctccagcc atcatgccgt tcaaagtgca ggacctttgg aacaggcagc
     6721 tttccttcca gccatagcat catgtccttt tcccgttcca catcataggt ggtcccttta
     6781 taccggctgt ccgtcatttt taaatatagg ttttcatttt ctcccaccag cttatatacc
     6841 ttagcaggag acattccttc cgtatctttt acgcagcggt atttttcgat cagttttttc
     6901 aattccggtg atattctcat tttagccata ctcttccttt ttcaatatta ttgaagcatt
     6961 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7021 ataggggttc cgcgcactcg gagccttgat gtgggcgctt tttgcagctg ttcggctgtg
     7081 cgctggccag acagttatgc acaggccagg cgggttttaa gagttttaat aagttttaaa
     7141 gagttttagg cggaaaaatc gccttttttc tcttttatat cagtcactta catgtgtgac
     7201 cggttcccaa tgtacggctt tgggttccca atgtacgggt tccggttccc aatgtacggc
     7261 tttgggttcc caatgtacgt gctatccaca ggaaagaagc tttttcgacc tttttcccct
     7321 gctagggcaa tttgccctag catctgctcc gtacattagg aaccggcgga tgcttcgccc
     7381 tcgatcaggt tgcggtagcg catgactagg atcgggccag cctgccccgc ctcctccttc
     7441 aaatcgtact ccggcaggtc atttgacccg atcagcttgc gcacggtgaa acagaacttc
     7501 ttgaactctc cggcgctgcc actgcgttcg tagatcgtct tgaacaacca tctggcttct
     7561 gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc gggatcgatc
     7621 aaaaagtaat cggggtgaac cgtcagcacg tccgggttct tgccttctgt gatctcgcgg
     7681 tacatccaat cagctagctc gatctcgatg tactccggcc gcccggtttc gctctttacg
     7741 atcttgtagc ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg gccgttcttg
     7801 gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta accgaatgca ggtttctacc
     7861 aggtcgtctt tctgctttcc gccatcggct cgccggcaga acttgagtac gtccgcaacg
     7921 tgtggacgga acacgcggcc gggcttgtct cccttccctt cccggtatcg gttcatggat
     7981 tcggttagat gggaaaccgc catcagtacc aggtcgtaat cccacacact ggccatgccg
     8041 gccggccctg cggaaacctc tacgtgcccg tctggaagct cgtagcggat cacctcgcca
     8101 gctcgtcggt cacgcttcga cagacggaaa acggccacgt ccatgatgct gcgactatcg
     8161 cgggtgccca cgtcatagag catcggaacg aaaaaatctg gttgctcgtc gcccttgggc
     8221 ggcttcctaa tcgacgacgc accggctgcc ggcggttgcc gggattcttt gcggattcga
     8281 tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg
     8341 gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt
     8401 accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc
     8461 cattgcaggg ccggcagaca acccagccgc ttacgcctgg ccaaccgccc gttcctccac
     8521 acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt
     8581 tagccgctaa aat
//
