LOCUS       sequence-342202-        8503 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 8503)
  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 8503)
  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_09216
FEATURES             Location/Qualifiers
     source          1..8503
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    703..727
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(794..1501)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(1601..1612)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     promoter        complement(2019..2364)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2056..2076)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3397..3649)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4249..4431)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4285..4463)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4626..4650
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4786..5374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4866..4885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5545..6339)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6340..6444)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6506..6700
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6766..7839)
                     /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(8268..8503,1..394))
                     /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 ttccatccgt gacctcaatg cgctgcttaa ccagctccac caggtcggcg gtggcccata
       61 tgtcgtaagg gcttggctgc accggaatca gcacgaagtc ggctgccttg atcgcggaca
      121 cagccaagtc cgccgcctgg ggcgctccgt cgatcactac gaagtcgcgc cggccgatgg
      181 ccttcacgtc gcggtcaatc gtcgggcggt cgatgccgac aacggttagc ggttgatctt
      241 cccgcacggc cgcccaatcg cgggcactgc cctggggatc ggaatcgact aacagaacat
      301 cggccccggc gagttgcagg gcgcgggcta gatgggttgc gatggtcgtc ttgcctgacc
      361 cgcctttctg gttaagtaca gcgataacct tcatgcgttc cccttgcgta tttgtttatt
      421 tactcatcgc atcatatacg cagcgaccgc atgacgcaag ctgttttact caaatacaca
      481 tcaccttttt agacggcggc gctcggtttc ttcagcggcc aagctggccg gccaggccgc
      541 cagcttggca tcagacaaac cggccaggat ttcatgcagc cgcacggctg actcccgagg
      601 ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct
      661 tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat
      721 cctgtcaaac actgatagtt taaacggatc acttcgtgtc cctcggtcac atgtgcatcc
      781 tctagtagcg gtcctgctga gcctcgacat gttgtcgcaa aattcgccct ggacccgccc
      841 aacgatttgt cgtcactgtc aaggtttgac ctgcacttca tttggggccc acatacacca
      901 aaaaaatgct gcataattct cggggcagca agtcggttac ccggccgccg tgctggaccg
      961 ggttgaatgg tgcccgtaac tttcggtaga gcggacggcc aatactcaac ttcaaggaat
     1021 ctcacccatg cgcgccggcg gggaaccgga gttcccttca gtgagcgtta ttagttcgcc
     1081 gctcggtgtg tcgtagatac tagcccctgg ggcacttttg aaatttgaat aagatttatg
     1141 taatcagtct tttaggtttg accggttctg ccgctttttt taaaattgga tttgtaataa
     1201 taaaacgcaa ttgtttgtta ttgtggcgct ctatcataga tgtcgctata aacctattca
     1261 gcacaatata ttgttttcat tttaatattg tacatataag tagtagggta caatcagtaa
     1321 attgaacgga gaatattatt cataaaaata cgatagtaac gggtgatata ttcattagaa
     1381 tgaaccgaaa ccggcggtaa ggatctgagc tacacatgct caggtttttt acaacgtgca
     1441 caacagaatt gaaagcaaat atcatgcgat cataggcttc tcgcatatct cattaaagca
     1501 ggacaagctc atgctgatcc agttgtagta gttgtctcag cagtagatga cttcctgtcg
     1561 aacaaagcct tgtaagctcc gtagatagca agacctccga ttgtagctcc caaagtgatc
     1621 ttagcgaact tcctcaacct aggccaagac ttagggttct tctgcacagt gtcagccatc
     1681 atagctgaga tcttagcgat tccctcgtca gacatctttg ctgactcctt tgcagcagcg
     1741 aaagcagtga tagcctgcct cacagccacc tcgtcaccgt cagaagccct gatagcgttc
     1801 acagccttag tcaaagctgt gtttgtctta agcttctcca agaatcctcc agcaagctta
     1861 gccacgttgt ccttgttcac tcccacagag ttgatagctg gcacagcagg cactgcgtgc
     1921 tcagccattg tatcgataat tgtaaatgta attgtaatgt tgtttgttgt ttgttgttgt
     1981 tggtaattgt tgtaaaaatg agctcttata ctcgagcgtg tcctctccaa atgaaatgaa
     2041 cttccttata tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc atcccttacg
     2101 tcagtggaga tgtcacatca atccacttgc tttgtagacg tggttggaac ctcttctttt
     2161 tccacgatgc tcctcgtggg tgggggtcca tctttgggac cactgtcggc agagagatct
     2221 tgaatgatag cctttccttt atcgcaatga tggcatttgt aggagccacc ttccttttct
     2281 actgtccttt cgatgaagtg acagatagct gggcaatgga atccgaggag gtttcccgaa
     2341 attatccttt gttgaaaagt ctcaatagcc ctttgatctt ctgagactgt atctttgaca
     2401 tttttggagt agaccagagt gtcgtgctcc accatgttga cgaagatttt cttcttgtca
     2461 ttgagtcgta aaagactctg tatgaactgt tcgccagtct tgacggcgag ttctgttaga
     2521 tcctcgattt gaatcttaga ctccatgcat ggccttagat tcagtaggaa ctaccttttt
     2581 agagactcca atctctatta cttgccttgg tttatgaagc aagccttgaa tcgtccatac
     2641 tggaatagta cttctgatct tgagaaatat gtctttctct gtgttcttga tgcaattagt
     2701 cctgaatctt ttgactgcat ctttaacctt cttgggaagg tatttgatct cctggagatt
     2761 gttactcggg tagatcgtct tgatgagact tgctgcgtag gcctctctaa ccatctgtgg
     2821 gtcagcattc tttctgaaat tgaagaggct aaccttctca ttatcagtgg tgaacatagt
     2881 gtcgtcacct tcaccttcga acttccttcc tagatcgtaa agatagagga aatcgtccat
     2941 tgtaatctcc ggggcaaagg agatctcttt tggggctgga tcactgctgg gccttttggt
     3001 tcctagcgtg agccagtggg ctttttgctt tggtgggctt gttagggcct tagcaaagct
     3061 cttgggcttg agttgagctt ctcctttggg gatgaagttc aacctgtctg tttgctgact
     3121 tgttgtgtac gcgtcagctg ctgctcttgc ctctgtaata gtggcaaatt tcttgtgtgc
     3181 aactccggga acgccgtttg ttgccgcctt tgtacaaccc cagtcatcgt atataccggc
     3241 atgtggaccg ttatacacaa cgtagtagtt gatatgaggg tgttgaatac ccgattctgc
     3301 tctgagagga gcaactgtgc tgttaagctc agatttttgt gggattggaa ttcctccaag
     3361 cttgaattct tgcagcgctc caattcagca agcgtcgatc tagtaacata gatgacaccg
     3421 cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg tattaaatgt
     3481 ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg cattacatgt
     3541 taattattac atgcttaacg taattcaaca gaaattatat gataatcatc gcaagaccgg
     3601 caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct gcttgactct
     3661 agatccagag tcccgctcaa atctcagtca caggaagaac aggtctagga ggaacaggaa
     3721 gagagaaatc aagctgccag aatcccacat catgccaatt tccatgtttg aaaccagcag
     3781 ctctaagcat tcccctagga gcataaccaa gagcctcatg catcctcaca gaaggatcat
     3841 taggaagtcc aatcacagca accacagact tgaaaccctg agcctccaaa gacttcaaaa
     3901 gatgagtgta aagagtagat ccaagaccag ttctttgatg cctaggagac acgtaaacag
     3961 tagactcagc agtccaatcg taagcattcc tagccttcca aggaccagca taagcaatac
     4021 cagccacttc accatccact tcagcaacaa gccaaggata cctctctcta agcctcacaa
     4081 gatcatcagt ccactcttgt ggttcttgag gctcagtcct gaaattcaca gtagaagtct
     4141 caatgtaatg attcacaata gtgcacacag caggcatatc agcctcagta gcccttctaa
     4201 tatcagcagg ccttctttca ggagacatgc gaaacgatcc agatccggtg cagattattt
     4261 ggattgagag tgaatatgag actctaattg gataccgagg ggaatttatg gaacgtcagt
     4321 ggagcatttt gacaagaaat atttgctagc tgatagtgac cttaggcgac ttttgaacgc
     4381 gcaataatgg tttctgacgt atgtgcttag ctcattaaac tccagaaacc cgcggctgag
     4441 tggctccttc aacgttgcgg ttctgtcagt tccaaacgta aaacggcttg tcccgcgtca
     4501 tcggcggggg tcataacgtg actcccttaa ttctccgctc cggatccgaa ttcggcacag
     4561 agtgttcaac cccagtacat taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc
     4621 aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc cgaccggcag
     4681 ctcggcacaa aatcaccact cgatacaggc agcccatcaa ccatgactac tgtccggtcc
     4741 aacagatccg agcaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4801 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4861 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4921 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4981 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5041 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5101 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5161 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5221 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5281 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5341 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5401 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5461 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5521 atatgagtaa acttggtctg acagctaaaa caattcatcc agtaaaatat aatattttat
     5581 tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat agctcgacat actgttcttc
     5641 cccgatatcc tccctgagcg accggacgca gaaggcaatg tcataccact tgtccgccct
     5701 gccgcttctc ccaagatcaa taaagccact tactttgcca tctttcacaa agatgttgct
     5761 gtctcccagg tcgccgtggg aaaagacaag ttcctcttcg ggcttttccg tctttaaaaa
     5821 atcatacagc tcgcgcggat ctttaaatgg agtgtcctct tcccagtttt cgcaatccac
     5881 atcggccaga tcgttattca gtaagtaatc caattcggct aagcggctgt ctaagctatt
     5941 cgtataggga caatccgata tgtcgatgga gtgaaagagc ctgatgcact ccgcatacag
     6001 ctcgataatc ttttcagggc tttgttcatc ttcatactct tccgagcaaa ggacgccatc
     6061 ggcctcactc atgagcagat tgctccagcc atcatgccgt tcaaagtgca ggacctttgg
     6121 aacaggcagc tttccttcca gccatagcat catgtccttt tcccgttcca catcataggt
     6181 ggtcccttta taccggctgt ccgtcatttt taaatatagg ttttcatttt ctcccaccag
     6241 cttatatacc ttagcaggag acattccttc cgtatctttt acgcagcggt atttttcgat
     6301 cagttttttc aattccggtg atattctcat tttagccata ctcttccttt ttcaatatta
     6361 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6421 aaataaacaa ataggggttc cgcgcactcg gagccttgat gtgggcgctt tttgcagctg
     6481 ttcggctgtg cgctggccag acagttatgc acaggccagg cgggttttaa gagttttaat
     6541 aagttttaaa gagttttagg cggaaaaatc gccttttttc tcttttatat cagtcactta
     6601 catgtgtgac cggttcccaa tgtacggctt tgggttccca atgtacgggt tccggttccc
     6661 aatgtacggc tttgggttcc caatgtacgt gctatccaca ggaaagaagc tttttcgacc
     6721 tttttcccct gctagggcaa tttgccctag catctgctcc gtacattagg aaccggcgga
     6781 tgcttcgccc tcgatcaggt tgcggtagcg catgactagg atcgggccag cctgccccgc
     6841 ctcctccttc aaatcgtact ccggcaggtc atttgacccg atcagcttgc gcacggtgaa
     6901 acagaacttc ttgaactctc cggcgctgcc actgcgttcg tagatcgtct tgaacaacca
     6961 tctggcttct gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc
     7021 gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg tccgggttct tgccttctgt
     7081 gatctcgcgg tacatccaat cagctagctc gatctcgatg tactccggcc gcccggtttc
     7141 gctctttacg atcttgtagc ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg
     7201 gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta accgaatgca
     7261 ggtttctacc aggtcgtctt tctgctttcc gccatcggct cgccggcaga acttgagtac
     7321 gtccgcaacg tgtggacgga acacgcggcc gggcttgtct cccttccctt cccggtatcg
     7381 gttcatggat tcggttagat gggaaaccgc catcagtacc aggtcgtaat cccacacact
     7441 ggccatgccg gccggccctg cggaaacctc tacgtgcccg tctggaagct cgtagcggat
     7501 cacctcgcca gctcgtcggt cacgcttcga cagacggaaa acggccacgt ccatgatgct
     7561 gcgactatcg cgggtgccca cgtcatagag catcggaacg aaaaaatctg gttgctcgtc
     7621 gcccttgggc ggcttcctaa tcgacgacgc accggctgcc ggcggttgcc gggattcttt
     7681 gcggattcga tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg
     7741 ttgccgctgg gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc
     7801 gccgatttgt accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg
     7861 gttccagtgc cattgcaggg ccggcagaca acccagccgc ttacgcctgg ccaaccgccc
     7921 gttcctccac acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg
     7981 ccgcctcctt tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg
     8041 tagctgcgcg atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta
     8101 catcttcagc ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg
     8161 cgtgtctgcc aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc
     8221 acttagcgtg tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt
     8281 gatttaattt cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat
     8341 tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg
     8401 gactcaagaa tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc
     8461 gtgcctttga tcgcccgcga cacgacaaag gccgcttgta gcc
//
