LOCUS       sequence-342330-        8362 bp    DNA     circular SYN 15-APR-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 8362)
  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 8362)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_12737
FEATURES             Location/Qualifiers
     source          1..8362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    641..665
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(732..1439)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1816..2161)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1853..1873)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3194..3446)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4046..4228)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4082..4260)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4423..4447
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4583..5171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4663..4682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5342..6136)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6137..6241)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6303..6497
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6563..7636)
                     /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(8065..8362,1..332))
                     /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 tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg ctgccttgat cgcggacaca
       61 gccaagtccg ccgcctgggg cgctccgtcg atcactacga agtcgcgccg gccgatggcc
      121 ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg ttgatcttcc
      181 cgcacggccg cccaatcgcg ggcactgccc tggggatcgg aatcgactaa cagaacatcg
      241 gccccggcga gttgcagggc gcgggctaga tgggttgcga tggtcgtctt gcctgacccg
      301 cctttctggt taagtacagc gataaccttc atgcgttccc cttgcgtatt tgtttattta
      361 ctcatcgcat catatacgca gcgaccgcat gacgcaagct gttttactca aatacacatc
      421 acctttttag acggcggcgc tcggtttctt cagcggccaa gctggccggc caggccgcca
      481 gcttggcatc agacaaaccg gccaggattt catgcagccg cacggctgac tcccgagggg
      541 aaccctgtgg ttggcatgca catacaaatg gacgaacgga taaacctttt cacgcccttt
      601 taaatatccg attattctaa taaacgctct tttctcttag gtttacccgc caatatatcc
      661 tgtcaaacac tgatagttta aacggatcac ttcgtgtccc tcggtcacat gtgcatcctc
      721 tagtagcggt cctgctgagc ctcgacatgt tgtcgcaaaa ttcgccctgg acccgcccaa
      781 cgatttgtcg tcactgtcaa ggtttgacct gcacttcatt tggggcccac atacaccaaa
      841 aaaatgctgc ataattctcg gggcagcaag tcggttaccc ggccgccgtg ctggaccggg
      901 ttgaatggtg cccgtaactt tcggtagagc ggacggccaa tactcaactt caaggaatct
      961 cacccatgcg cgccggcggg gaaccggagt tcccttcagt gagcgttatt agttcgccgc
     1021 tcggtgtgtc gtagatacta gcccctgggg cacttttgaa atttgaataa gatttatgta
     1081 atcagtcttt taggtttgac cggttctgcc gcttttttta aaattggatt tgtaataata
     1141 aaacgcaatt gtttgttatt gtggcgctct atcatagatg tcgctataaa cctattcagc
     1201 acaatatatt gttttcattt taatattgta catataagta gtagggtaca atcagtaaat
     1261 tgaacggaga atattattca taaaaatacg atagtaacgg gtgatatatt cattagaatg
     1321 aaccgaaacc ggcggtaagg atctgagcta cacatgctca ggttttttac aacgtgcaca
     1381 acagaattga aagcaaatat catgcgatca taggcttctc gcatatctca ttaaagcagg
     1441 acaagctcag tggaagatct tccagtactc gtcgtagaac ttgatcaact cgtcagcgtc
     1501 agcgtccttc attcccttag gtggtgtgtt cttgatagcg tccctcaaag tctcaggagt
     1561 gtatcccttc ttctccaagt cagtcaaagc gtacaaagac ttctcctcca tcttctctga
     1621 caacttcttc aactggttca aagacttagt cttaagcatc acctctcctc ccaccttctt
     1681 gaacttgttc accaagttag tcaacttctc aggcaccaat cccattgtat cgataattgt
     1741 aaatgtaatt gtaatgttgt ttgttgtttg ttgttgttgg taattgttgt aaaaatgagc
     1801 tcttatactc gagcgtgtcc tctccaaatg aaatgaactt ccttatatag aggaagggtc
     1861 ttgcgaagga tagtgggatt gtgcgtcatc ccttacgtca gtggagatgt cacatcaatc
     1921 cacttgcttt gtagacgtgg ttggaacctc ttctttttcc acgatgctcc tcgtgggtgg
     1981 gggtccatct ttgggaccac tgtcggcaga gagatcttga atgatagcct ttcctttatc
     2041 gcaatgatgg catttgtagg agccaccttc cttttctact gtcctttcga tgaagtgaca
     2101 gatagctggg caatggaatc cgaggaggtt tcccgaaatt atcctttgtt gaaaagtctc
     2161 aatagccctt tgatcttctg agactgtatc tttgacattt ttggagtaga ccagagtgtc
     2221 gtgctccacc atgttgacga agattttctt cttgtcattg agtcgtaaaa gactctgtat
     2281 gaactgttcg ccagtcttga cggcgagttc tgttagatcc tcgatttgaa tcttagactc
     2341 catgcatggc cttagattca gtaggaacta cctttttaga gactccaatc tctattactt
     2401 gccttggttt atgaagcaag ccttgaatcg tccatactgg aatagtactt ctgatcttga
     2461 gaaatatgtc tttctctgtg ttcttgatgc aattagtcct gaatcttttg actgcatctt
     2521 taaccttctt gggaaggtat ttgatctcct ggagattgtt actcgggtag atcgtcttga
     2581 tgagacttgc tgcgtaggcc tctctaacca tctgtgggtc agcattcttt ctgaaattga
     2641 agaggctaac cttctcatta tcagtggtga acatagtgtc gtcaccttca ccttcgaact
     2701 tccttcctag atcgtaaaga tagaggaaat cgtccattgt aatctccggg gcaaaggaga
     2761 tctcttttgg ggctggatca ctgctgggcc ttttggttcc tagcgtgagc cagtgggctt
     2821 tttgctttgg tgggcttgtt agggccttag caaagctctt gggcttgagt tgagcttctc
     2881 ctttggggat gaagttcaac ctgtctgttt gctgacttgt tgtgtacgcg tcagctgctg
     2941 ctcttgcctc tgtaatagtg gcaaatttct tgtgtgcaac tccgggaacg ccgtttgttg
     3001 ccgcctttgt acaaccccag tcatcgtata taccggcatg tggaccgtta tacacaacgt
     3061 agtagttgat atgagggtgt tgaatacccg attctgctct gagaggagca actgtgctgt
     3121 taagctcaga tttttgtggg attggaattc ctccaagctt gaattcttgc agcgctccaa
     3181 ttcagcaagc gtcgatctag taacatagat gacaccgcgc gcgataattt atcctagttt
     3241 gcgcgctata ttttgttttc tatcgcgtat taaatgtata attgcgggac tctaatcata
     3301 aaaacccatc tcataaataa cgtcatgcat tacatgttaa ttattacatg cttaacgtaa
     3361 ttcaacagaa attatatgat aatcatcgca agaccggcaa caggattcaa tcttaagaaa
     3421 ctttattgcc aaatgtttga acgatctgct tgactctaga tccagagtcc cgctcaaatc
     3481 tcagtcacag gaagaacagg tctaggagga acaggaagag agaaatcaag ctgccagaat
     3541 cccacatcat gccaatttcc atgtttgaaa ccagcagctc taagcattcc cctaggagca
     3601 taaccaagag cctcatgcat cctcacagaa ggatcattag gaagtccaat cacagcaacc
     3661 acagacttga aaccctgagc ctccaaagac ttcaaaagat gagtgtaaag agtagatcca
     3721 agaccagttc tttgatgcct aggagacacg taaacagtag actcagcagt ccaatcgtaa
     3781 gcattcctag ccttccaagg accagcataa gcaataccag ccacttcacc atccacttca
     3841 gcaacaagcc aaggatacct ctctctaagc ctcacaagat catcagtcca ctcttgtggt
     3901 tcttgaggct cagtcctgaa attcacagta gaagtctcaa tgtaatgatt cacaatagtg
     3961 cacacagcag gcatatcagc ctcagtagcc cttctaatat cagcaggcct tctttcagga
     4021 gacatgcgaa acgatccaga tccggtgcag attatttgga ttgagagtga atatgagact
     4081 ctaattggat accgagggga atttatggaa cgtcagtgga gcattttgac aagaaatatt
     4141 tgctagctga tagtgacctt aggcgacttt tgaacgcgca ataatggttt ctgacgtatg
     4201 tgcttagctc attaaactcc agaaacccgc ggctgagtgg ctccttcaac gttgcggttc
     4261 tgtcagttcc aaacgtaaaa cggcttgtcc cgcgtcatcg gcgggggtca taacgtgact
     4321 cccttaattc tccgctccgg atccgaattc ggcacagagt gttcaacccc agtacattaa
     4381 aaacgtccgc aatgtgttat taagttgtct aagcgtcaat ttgtttacac cacaatatat
     4441 cctgccacca gccagccaac agctccccga ccggcagctc ggcacaaaat caccactcga
     4501 tacaggcagc ccatcaacca tgactactgt ccggtccaac agatccgagc aggaaccgta
     4561 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4621 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4741 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4801 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4861 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4921 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4981 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     5041 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     5101 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     5161 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     5221 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     5281 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     5341 gctaaaacaa ttcatccagt aaaatataat attttatttt ctcccaatca ggcttgatcc
     5401 ccagtaagtc aaaaaatagc tcgacatact gttcttcccc gatatcctcc ctgagcgacc
     5461 ggacgcagaa ggcaatgtca taccacttgt ccgccctgcc gcttctccca agatcaataa
     5521 agccacttac tttgccatct ttcacaaaga tgttgctgtc tcccaggtcg ccgtgggaaa
     5581 agacaagttc ctcttcgggc ttttccgtct ttaaaaaatc atacagctcg cgcggatctt
     5641 taaatggagt gtcctcttcc cagttttcgc aatccacatc ggccagatcg ttattcagta
     5701 agtaatccaa ttcggctaag cggctgtcta agctattcgt atagggacaa tccgatatgt
     5761 cgatggagtg aaagagcctg atgcactccg catacagctc gataatcttt tcagggcttt
     5821 gttcatcttc atactcttcc gagcaaagga cgccatcggc ctcactcatg agcagattgc
     5881 tccagccatc atgccgttca aagtgcagga cctttggaac aggcagcttt ccttccagcc
     5941 atagcatcat gtccttttcc cgttccacat cataggtggt ccctttatac cggctgtccg
     6001 tcatttttaa atataggttt tcattttctc ccaccagctt atatacctta gcaggagaca
     6061 ttccttccgt atcttttacg cagcggtatt tttcgatcag ttttttcaat tccggtgata
     6121 ttctcatttt agccatactc ttcctttttc aatattattg aagcatttat cagggttatt
     6181 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6241 gcactcggag ccttgatgtg ggcgcttttt gcagctgttc ggctgtgcgc tggccagaca
     6301 gttatgcaca ggccaggcgg gttttaagag ttttaataag ttttaaagag ttttaggcgg
     6361 aaaaatcgcc ttttttctct tttatatcag tcacttacat gtgtgaccgg ttcccaatgt
     6421 acggctttgg gttcccaatg tacgggttcc ggttcccaat gtacggcttt gggttcccaa
     6481 tgtacgtgct atccacagga aagaagcttt ttcgaccttt ttcccctgct agggcaattt
     6541 gccctagcat ctgctccgta cattaggaac cggcggatgc ttcgccctcg atcaggttgc
     6601 ggtagcgcat gactaggatc gggccagcct gccccgcctc ctccttcaaa tcgtactccg
     6661 gcaggtcatt tgacccgatc agcttgcgca cggtgaaaca gaacttcttg aactctccgg
     6721 cgctgccact gcgttcgtag atcgtcttga acaaccatct ggcttctgcc ttgcctgcgg
     6781 cgcggcgtgc caggcggtag agaaaacggc cgatgccggg atcgatcaaa aagtaatcgg
     6841 ggtgaaccgt cagcacgtcc gggttcttgc cttctgtgat ctcgcggtac atccaatcag
     6901 ctagctcgat ctcgatgtac tccggccgcc cggtttcgct ctttacgatc ttgtagcggc
     6961 taatcaaggc ttcaccctcg gataccgtca ccaggcggcc gttcttggcc ttcttcgtac
     7021 gctgcatggc aacgtgcgtg gtgtttaacc gaatgcaggt ttctaccagg tcgtctttct
     7081 gctttccgcc atcggctcgc cggcagaact tgagtacgtc cgcaacgtgt ggacggaaca
     7141 cgcggccggg cttgtctccc ttcccttccc ggtatcggtt catggattcg gttagatggg
     7201 aaaccgccat cagtaccagg tcgtaatccc acacactggc catgccggcc ggccctgcgg
     7261 aaacctctac gtgcccgtct ggaagctcgt agcggatcac ctcgccagct cgtcggtcac
     7321 gcttcgacag acggaaaacg gccacgtcca tgatgctgcg actatcgcgg gtgcccacgt
     7381 catagagcat cggaacgaaa aaatctggtt gctcgtcgcc cttgggcggc ttcctaatcg
     7441 acgacgcacc ggctgccggc ggttgccggg attctttgcg gattcgatca gcggccgctt
     7501 gccacgattc accggggcgt gcttctgcct cgatgcgttg ccgctgggcg gcctgcgcgg
     7561 ccttcaactt ctccaccagg tcatcaccca gcgccgcgcc gatttgtacc gggccggatg
     7621 gtttgcgacc gctcacgccg attcctcggg cttgggggtt ccagtgccat tgcagggccg
     7681 gcagacaacc cagccgctta cgcctggcca accgcccgtt cctccacaca tggggcattc
     7741 cacggcgtcg gtgcctggtt gttcttgatt ttccatgccg cctcctttag ccgctaaaat
     7801 tcatctactc atttattcat ttgctcattt actctggtag ctgcgcgatg tattcagata
     7861 gcagctcggt aatggtcttg ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct
     7921 ccgccggcaa ctgaaagttg acccgcttca tggctggcgt gtctgccagg ctggccaacg
     7981 ttgcagcctt gctgctgcgt gcgctcggac ggccggcact tagcgtgttt gtgcttttgc
     8041 tcattttctc tttacctcat taactcaaat gagttttgat ttaatttcag cggccagcgc
     8101 ctggacctcg cgggcagcgt cgccctcggg ttctgattca agaacggttg tgccggcggc
     8161 ggcagtgcct gggtagctca cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta
     8221 cccggccagc gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac
     8281 gacaaaggcc gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa ccagctccac
     8341 caggtcggcg gtggcccata tg
//
