LOCUS       sequence-342545-        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_15127
FEATURES             Location/Qualifiers
     source          1..9484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(240..869)
                     /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    1178..1202
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1269..1976)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3475..3820)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3512..3532)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4853..5105)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5705..5887)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5741..5919)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6082..6106
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6242..6830)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6322..6341)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7001..7795)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7796..7900)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7962..8156
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8222..9295)
                     /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 catttactct ggtagctgcg cgatgtattc agatagcagc tcggtaatgg tcttgccttg
       61 gcgtaccgcg tacatcttca gcttggtgtg atcctccgcc ggcaactgaa agttgacccg
      121 cttcatggct ggcgtgtctg ccaggctggc caacgttgca gccttgctgc tgcgtgcgct
      181 cggacggccg gcacttagcg tgtttgtgct tttgctcatt ttctctttac ctcattaact
      241 caaatgagtt ttgatttaat ttcagcggcc agcgcctgga cctcgcgggc agcgtcgccc
      301 tcgggttctg attcaagaac ggttgtgccg gcggcggcag tgcctgggta gctcacgcgc
      361 tgcgtgatac gggactcaag aatgggcagc tcgtacccgg ccagcgcctc ggcaacctca
      421 ccgccgatgc gcgtgccttt gatcgcccgc gacacgacaa aggccgcttg tagccttcca
      481 tccgtgacct caatgcgctg cttaaccagc tccaccaggt cggcggtggc ccatatgtcg
      541 taagggcttg gctgcaccgg aatcagcacg aagtcggctg ccttgatcgc ggacacagcc
      601 aagtccgccg cctggggcgc tccgtcgatc actacgaagt cgcgccggcc gatggccttc
      661 acgtcgcggt caatcgtcgg gcggtcgatg ccgacaacgg ttagcggttg atcttcccgc
      721 acggccgccc aatcgcgggc actgccctgg ggatcggaat cgactaacag aacatcggcc
      781 ccggcgagtt gcagggcgcg ggctagatgg gttgcgatgg tcgtcttgcc tgacccgcct
      841 ttctggttaa gtacagcgat aaccttcatg cgttcccctt gcgtatttgt ttatttactc
      901 atcgcatcat atacgcagcg accgcatgac gcaagctgtt ttactcaaat acacatcacc
      961 tttttagacg gcggcgctcg gtttcttcag cggccaagct ggccggccag gccgccagct
     1021 tggcatcaga caaaccggcc aggatttcat gcagccgcac ggctgactcc cgaggggaac
     1081 cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac gcccttttaa
     1141 atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa tatatcctgt
     1201 caaacactga tagtttaaac ggatcacttc gtgtccctcg gtcacatgtg catcctctag
     1261 tagcggtcct gctgagcctc gacatgttgt cgcaaaattc gccctggacc cgcccaacga
     1321 tttgtcgtca ctgtcaaggt ttgacctgca cttcatttgg ggcccacata caccaaaaaa
     1381 atgctgcata attctcgggg cagcaagtcg gttacccggc cgccgtgctg gaccgggttg
     1441 aatggtgccc gtaactttcg gtagagcgga cggccaatac tcaacttcaa ggaatctcac
     1501 ccatgcgcgc cggcggggaa ccggagttcc cttcagtgag cgttattagt tcgccgctcg
     1561 gtgtgtcgta gatactagcc cctggggcac ttttgaaatt tgaataagat ttatgtaatc
     1621 agtcttttag gtttgaccgg ttctgccgct ttttttaaaa ttggatttgt aataataaaa
     1681 cgcaattgtt tgttattgtg gcgctctatc atagatgtcg ctataaacct attcagcaca
     1741 atatattgtt ttcattttaa tattgtacat ataagtagta gggtacaatc agtaaattga
     1801 acggagaata ttattcataa aaatacgata gtaacgggtg atatattcat tagaatgaac
     1861 cgaaaccggc ggtaaggatc tgagctacac atgctcaggt tttttacaac gtgcacaaca
     1921 gaattgaaag caaatatcat gcgatcatag gcttctcgca tatctcatta aagcaggaca
     1981 agctcacctc cttggattaa ttgaagtgtc aatccttgga ggagccatgt gctccttgta
     2041 gtagtccttg taccttgcag cagtagccct ctcaaggtta gaagcagtgt tttcagatga
     2101 cctaaaagta gtaccgtaga tgtcagatgg ttttgtcttt ctcttgaacc acctaccaaa
     2161 caacattctt tggtactctg tagcgataga gatctcgtcc aagtctgcgt aaccgtttct
     2221 gatcaagtgc tccctagccc tagcagtctg gatagccaac tcgtcctctc ctccgaatcc
     2281 ctttgacaca gccctcaaca taccctcgtt gtacaacaag ttcttgttag aaagagttgc
     2341 caagtatcct gacaatgtgt caaattttgg gtttgtaggt gacaaaagat cgttgaaatg
     2401 gatatcaaga agcctaaaaa cagaattagc tgagattcct ccctccttcc acttaaggaa
     2461 ctgctgcctc tgcaatgtga taaccaagtt catagttgca gcgtcgtcag gcttagcgaa
     2521 tgtcaatgca gttgccactg ccttgttacc gtactgtgcc acgaatgagt cgatcaaaga
     2581 agccttagag ttagggtttt tagcattgta catagaaata aatgcaccaa atgtgtgaac
     2641 atttctgtcc aacaaagcgt ccaatcccct gttaagcctc aacttcttgt agatatgctt
     2701 tggcctcaac aagtcaactc taacccactt gttaatcaat gctgactgaa gagccaatgc
     2761 aagctcccta gtcttaacgt cctcccttgc cctagccaag attgttgcca actgctcctt
     2821 tcctccgaaa cccttagtaa gtgtcttaag aagagaatct tttgttgact tagttgagat
     2881 aacaagttta atataatcgt caagcacctc catcttaggt gaagccaaag ccaagtatcc
     2941 gtcagacctc aacttcaaaa gagtgaaaac gtcgtccact gacttcctct tctgcaacca
     3001 tgaaagaagc tgtgcgttcc tcaacctctg gatagtgttc atctcaccac cagccctttg
     3061 tgcagtcatc aaagcgtttg ccacagcgtc atcaccgtag tgtgccatga acataccgat
     3121 cactgaagcc ttgttgttat tgttctgtgt attcaactcc ttcacatagt tttcaagtgc
     3181 cttaaggttt ggtgaagcca atgcgtcgtc cattccattt gtcaacttga acctcctaag
     3241 gattggctct gctggtgggt ccttagagat ccatgatccc aagttgatgt tagacacacc
     3301 tgccttagcc aagtctgtca acttctccaa tccagctgag atcaccctgt tctcaccgtt
     3361 ctcagtattc tgagtagaca ccattgtatc gataattgta aatgtaattg taatgttgtt
     3421 tgttgtttgt tgttgttggt aattgttgta aaaatgagct cttatactcg agcgtgtcct
     3481 ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg
     3541 tgcgtcatcc cttacgtcag tggagatgtc acatcaatcc acttgctttg tagacgtggt
     3601 tggaacctct tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact
     3661 gtcggcagag agatcttgaa tgatagcctt tcctttatcg caatgatggc atttgtagga
     3721 gccaccttcc ttttctactg tcctttcgat gaagtgacag atagctgggc aatggaatcc
     3781 gaggaggttt cccgaaatta tcctttgttg aaaagtctca atagcccttt gatcttctga
     3841 gactgtatct ttgacatttt tggagtagac cagagtgtcg tgctccacca tgttgacgaa
     3901 gattttcttc ttgtcattga gtcgtaaaag actctgtatg aactgttcgc cagtcttgac
     3961 ggcgagttct gttagatcct cgatttgaat cttagactcc atgcatggcc ttagattcag
     4021 taggaactac ctttttagag actccaatct ctattacttg ccttggttta tgaagcaagc
     4081 cttgaatcgt ccatactgga atagtacttc tgatcttgag aaatatgtct ttctctgtgt
     4141 tcttgatgca attagtcctg aatcttttga ctgcatcttt aaccttcttg ggaaggtatt
     4201 tgatctcctg gagattgtta ctcgggtaga tcgtcttgat gagacttgct gcgtaggcct
     4261 ctctaaccat ctgtgggtca gcattctttc tgaaattgaa gaggctaacc ttctcattat
     4321 cagtggtgaa catagtgtcg tcaccttcac cttcgaactt ccttcctaga tcgtaaagat
     4381 agaggaaatc gtccattgta atctccgggg caaaggagat ctcttttggg gctggatcac
     4441 tgctgggcct tttggttcct agcgtgagcc agtgggcttt ttgctttggt gggcttgtta
     4501 gggccttagc aaagctcttg ggcttgagtt gagcttctcc tttggggatg aagttcaacc
     4561 tgtctgtttg ctgacttgtt gtgtacgcgt cagctgctgc tcttgcctct gtaatagtgg
     4621 caaatttctt gtgtgcaact ccgggaacgc cgtttgttgc cgcctttgta caaccccagt
     4681 catcgtatat accggcatgt ggaccgttat acacaacgta gtagttgata tgagggtgtt
     4741 gaatacccga ttctgctctg agaggagcaa ctgtgctgtt aagctcagat ttttgtggga
     4801 ttggaattcc tccaagcttg aattcttgca gcgctccaat tcagcaagcg tcgatctagt
     4861 aacatagatg acaccgcgcg cgataattta tcctagtttg cgcgctatat tttgttttct
     4921 atcgcgtatt aaatgtataa ttgcgggact ctaatcataa aaacccatct cataaataac
     4981 gtcatgcatt acatgttaat tattacatgc ttaacgtaat tcaacagaaa ttatatgata
     5041 atcatcgcaa gaccggcaac aggattcaat cttaagaaac tttattgcca aatgtttgaa
     5101 cgatctgctt gactctagat ccagagtccc gctcaaatct cagtcacagg aagaacaggt
     5161 ctaggaggaa caggaagaga gaaatcaagc tgccagaatc ccacatcatg ccaatttcca
     5221 tgtttgaaac cagcagctct aagcattccc ctaggagcat aaccaagagc ctcatgcatc
     5281 ctcacagaag gatcattagg aagtccaatc acagcaacca cagacttgaa accctgagcc
     5341 tccaaagact tcaaaagatg agtgtaaaga gtagatccaa gaccagttct ttgatgccta
     5401 ggagacacgt aaacagtaga ctcagcagtc caatcgtaag cattcctagc cttccaagga
     5461 ccagcataag caataccagc cacttcacca tccacttcag caacaagcca aggatacctc
     5521 tctctaagcc tcacaagatc atcagtccac tcttgtggtt cttgaggctc agtcctgaaa
     5581 ttcacagtag aagtctcaat gtaatgattc acaatagtgc acacagcagg catatcagcc
     5641 tcagtagccc ttctaatatc agcaggcctt ctttcaggag acatgcgaaa cgatccagat
     5701 ccggtgcaga ttatttggat tgagagtgaa tatgagactc taattggata ccgaggggaa
     5761 tttatggaac gtcagtggag cattttgaca agaaatattt gctagctgat agtgacctta
     5821 ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt gcttagctca ttaaactcca
     5881 gaaacccgcg gctgagtggc tccttcaacg ttgcggttct gtcagttcca aacgtaaaac
     5941 ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc ccttaattct ccgctccgga
     6001 tccgaattcg gcacagagtg ttcaacccca gtacattaaa aacgtccgca atgtgttatt
     6061 aagttgtcta agcgtcaatt tgtttacacc acaatatatc ctgccaccag ccagccaaca
     6121 gctccccgac cggcagctcg gcacaaaatc accactcgat acaggcagcc catcaaccat
     6181 gactactgtc cggtccaaca gatccgagca ggaaccgtaa aaaggccgcg ttgctggcgt
     6241 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6301 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6361 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6421 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6481 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6541 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6601 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6661 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6721 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6781 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6841 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6901 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6961 aatcaatcta aagtatatat gagtaaactt ggtctgacag ctaaaacaat tcatccagta
     7021 aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca aaaaatagct
     7081 cgacatactg ttcttccccg atatcctccc tgagcgaccg gacgcagaag gcaatgtcat
     7141 accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact ttgccatctt
     7201 tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc tcttcgggct
     7261 tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg tcctcttccc
     7321 agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat tcggctaagc
     7381 ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga aagagcctga
     7441 tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca tactcttccg
     7501 agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca tgccgttcaa
     7561 agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg tccttttccc
     7621 gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa tataggtttt
     7681 cattttctcc caccagctta tataccttag caggagacat tccttccgta tcttttacgc
     7741 agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta gccatactct
     7801 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     7861 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cactcggagc cttgatgtgg
     7921 gcgctttttg cagctgttcg gctgtgcgct ggccagacag ttatgcacag gccaggcggg
     7981 ttttaagagt tttaataagt tttaaagagt tttaggcgga aaaatcgcct tttttctctt
     8041 ttatatcagt cacttacatg tgtgaccggt tcccaatgta cggctttggg ttcccaatgt
     8101 acgggttccg gttcccaatg tacggctttg ggttcccaat gtacgtgcta tccacaggaa
     8161 agaagctttt tcgacctttt tcccctgcta gggcaatttg ccctagcatc tgctccgtac
     8221 attaggaacc ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg actaggatcg
     8281 ggccagcctg ccccgcctcc tccttcaaat cgtactccgg caggtcattt gacccgatca
     8341 gcttgcgcac ggtgaaacag aacttcttga actctccggc gctgccactg cgttcgtaga
     8401 tcgtcttgaa caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga
     8461 gaaaacggcc gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg
     8521 ggttcttgcc ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc tcgatgtact
     8581 ccggccgccc ggtttcgctc tttacgatct tgtagcggct aatcaaggct tcaccctcgg
     8641 ataccgtcac caggcggccg ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg
     8701 tgtttaaccg aatgcaggtt tctaccaggt cgtctttctg ctttccgcca tcggctcgcc
     8761 ggcagaactt gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc ttgtctccct
     8821 tcccttcccg gtatcggttc atggattcgg ttagatggga aaccgccatc agtaccaggt
     8881 cgtaatccca cacactggcc atgccggccg gccctgcgga aacctctacg tgcccgtctg
     8941 gaagctcgta gcggatcacc tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg
     9001 ccacgtccat gatgctgcga ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa
     9061 aatctggttg ctcgtcgccc ttgggcggct tcctaatcga cgacgcaccg gctgccggcg
     9121 gttgccggga ttctttgcgg attcgatcag cggccgcttg ccacgattca ccggggcgtg
     9181 cttctgcctc gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt
     9241 catcacccag cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg ctcacgccga
     9301 ttcctcgggc ttgggggttc cagtgccatt gcagggccgg cagacaaccc agccgcttac
     9361 gcctggccaa ccgcccgttc ctccacacat ggggcattcc acggcgtcgg tgcctggttg
     9421 ttcttgattt tccatgccgc ctcctttagc cgctaaaatt catctactca tttattcatt
     9481 tgct
//
