LOCUS       sequence-342349-        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_15125a
FEATURES             Location/Qualifiers
     source          1..9484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(330..959)
                     /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    1268..1292
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1359..2066)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3565..3910)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3602..3622)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4943..5195)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5795..5977)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5831..6009)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6172..6196
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6332..6920)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6412..6431)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7091..7885)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7886..7990)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8052..8246
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8312..9385)
                     /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 cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct
       61 aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc
      121 agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg
      181 atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc
      241 caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct
      301 tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc
      361 agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg
      421 gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc
      481 tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc
      541 gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc
      601 tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg
      661 aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc
      721 actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg
      781 ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg
      841 ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg
      901 gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg
      961 cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg accgcatgac
     1021 gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag
     1081 cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat
     1141 gcagccgcac ggctgactcc cgaggggaac cctgtggttg gcatgcacat acaaatggac
     1201 gaacggataa accttttcac gcccttttaa atatccgatt attctaataa acgctctttt
     1261 ctcttaggtt tacccgccaa tatatcctgt caaacactga tagtttaaac ggatcacttc
     1321 gtgtccctcg gtcacatgtg catcctctag tagcggtcct gctgagcctc gacatgttgt
     1381 cgcaaaattc gccctggacc cgcccaacga tttgtcgtca ctgtcaaggt ttgacctgca
     1441 cttcatttgg ggcccacata caccaaaaaa atgctgcata attctcgggg cagcaagtcg
     1501 gttacccggc cgccgtgctg gaccgggttg aatggtgccc gtaactttcg gtagagcgga
     1561 cggccaatac tcaacttcaa ggaatctcac ccatgcgcgc cggcggggaa ccggagttcc
     1621 cttcagtgag cgttattagt tcgccgctcg gtgtgtcgta gatactagcc cctggggcac
     1681 ttttgaaatt tgaataagat ttatgtaatc agtcttttag gtttgaccgg ttctgccgct
     1741 ttttttaaaa ttggatttgt aataataaaa cgcaattgtt tgttattgtg gcgctctatc
     1801 atagatgtcg ctataaacct attcagcaca atatattgtt ttcattttaa tattgtacat
     1861 ataagtagta gggtacaatc agtaaattga acggagaata ttattcataa aaatacgata
     1921 gtaacgggtg atatattcat tagaatgaac cgaaaccggc ggtaaggatc tgagctacac
     1981 atgctcaggt tttttacaac gtgcacaaca gaattgaaag caaatatcat gcgatcatag
     2041 gcttctcgca tatctcatta aagcaggaca agctcacctc ctagggttga tagaagtgtc
     2101 gatccttggt ggtgccatgt gctctttgta gtagtccttg tacctagctg cagtagccct
     2161 ctccaagttt gaagcagtgt tctcagaaga tctgaagata gtaccgtaga tgtcagatgg
     2221 ctctgtctcc ctcttgaacc accttccgaa caacatcctc tggtattcag ttgcgataga
     2281 gatctcgtcc aagtcagcgt aaccgttcct gataaggtgc tccctagccc ttgcagtctg
     2341 gatagccaac tcgtcctcac caccgaaacc ctttgacaca gccctcaaca ttccctcgtt
     2401 gtacaacaag ttcttgttag acaaagtagc caagtatcct gacaaagtgt cgaacttagg
     2461 attagtagga gacaaaaggt cgttgaagtt gatgtcaaga agcctgaaca cagagtttgc
     2521 tgagattcca ccctccttcc acttcaagaa ctgctgcctc tgcaatgtga tcaccaagtt
     2581 catagttgca gcgtcgtctg gcttagcgaa agtcaaagca gttgccactg ctttatttcc
     2641 gtactgagcc acaaatgagt cgatcaaaga tgcctttgaa tttgggtttt ttgcgttata
     2701 cattgagatg aaagctccga aagtgtgcac gttcctgtca agcaatgcgt ccaatcccct
     2761 gttcaacctc aactttttgt agatatgttt aggtctaaga agctccaccc tcacccactt
     2821 gttgataaga gcgttctgca aagctgaagc aagctccctt gtcttaacgt cctcccttgc
     2881 tctagcaagg attgttgcaa gctgctcctc tccaccaaat cctttagtca aagttttcaa
     2941 aagtgagtct ttagtagact ttgttgaagt aaccaattta atgtagtcgt caagcacttc
     3001 catcttagga gaagccaatg ccaaatatcc atcagacctc aatttaagca aagtaaaaac
     3061 gtcatcaact gatttcctct tctgcaacca agaaagcaac tgagcgtgcc taagcctctg
     3121 gatagtgttc atctcaccac cagccctctg tgcagtcatc aatgcgtttg ccacagcgtc
     3181 gtcaccgtag tgagccatga acataccgat cactgaagcc ttgttgttgt tgttctgagt
     3241 attcaactct ttaacataat tttccaatgc cttaaggtta ggtgaagcaa gagcgtcgtc
     3301 cattccgttt gtcaacttga acctcctcaa gataggctca gctggagggt ccttagagat
     3361 ccatgatcca aggttgatct tagacactcc agccttagca aggtctgtaa gcttctccaa
     3421 tcctgctgag atcaccctgt tctcaccatt ctcagtgttc tgtgtagaca ccattgtatc
     3481 gataattgta aatgtaattg taatgttgtt tgttgtttgt tgttgttggt aattgttgta
     3541 aaaatgagct cttatactcg agcgtgtcct ctccaaatga aatgaacttc cttatataga
     3601 ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatgtc
     3661 acatcaatcc acttgctttg tagacgtggt tggaacctct tctttttcca cgatgctcct
     3721 cgtgggtggg ggtccatctt tgggaccact gtcggcagag agatcttgaa tgatagcctt
     3781 tcctttatcg caatgatggc atttgtagga gccaccttcc ttttctactg tcctttcgat
     3841 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgaaatta tcctttgttg
     3901 aaaagtctca atagcccttt gatcttctga gactgtatct ttgacatttt tggagtagac
     3961 cagagtgtcg tgctccacca tgttgacgaa gattttcttc ttgtcattga gtcgtaaaag
     4021 actctgtatg aactgttcgc cagtcttgac ggcgagttct gttagatcct cgatttgaat
     4081 cttagactcc atgcatggcc ttagattcag taggaactac ctttttagag actccaatct
     4141 ctattacttg ccttggttta tgaagcaagc cttgaatcgt ccatactgga atagtacttc
     4201 tgatcttgag aaatatgtct ttctctgtgt tcttgatgca attagtcctg aatcttttga
     4261 ctgcatcttt aaccttcttg ggaaggtatt tgatctcctg gagattgtta ctcgggtaga
     4321 tcgtcttgat gagacttgct gcgtaggcct ctctaaccat ctgtgggtca gcattctttc
     4381 tgaaattgaa gaggctaacc ttctcattat cagtggtgaa catagtgtcg tcaccttcac
     4441 cttcgaactt ccttcctaga tcgtaaagat agaggaaatc gtccattgta atctccgggg
     4501 caaaggagat ctcttttggg gctggatcac tgctgggcct tttggttcct agcgtgagcc
     4561 agtgggcttt ttgctttggt gggcttgtta gggccttagc aaagctcttg ggcttgagtt
     4621 gagcttctcc tttggggatg aagttcaacc tgtctgtttg ctgacttgtt gtgtacgcgt
     4681 cagctgctgc tcttgcctct gtaatagtgg caaatttctt gtgtgcaact ccgggaacgc
     4741 cgtttgttgc cgcctttgta caaccccagt catcgtatat accggcatgt ggaccgttat
     4801 acacaacgta gtagttgata tgagggtgtt gaatacccga ttctgctctg agaggagcaa
     4861 ctgtgctgtt aagctcagat ttttgtggga ttggaattcc tccaagcttg aattcttgca
     4921 gcgctccaat tcagcaagcg tcgatctagt aacatagatg acaccgcgcg cgataattta
     4981 tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact
     5041 ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc
     5101 ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat
     5161 cttaagaaac tttattgcca aatgtttgaa cgatctgctt gactctagat ccagagtccc
     5221 gctcaaatct cagtcacagg aagaacaggt ctaggaggaa caggaagaga gaaatcaagc
     5281 tgccagaatc ccacatcatg ccaatttcca tgtttgaaac cagcagctct aagcattccc
     5341 ctaggagcat aaccaagagc ctcatgcatc ctcacagaag gatcattagg aagtccaatc
     5401 acagcaacca cagacttgaa accctgagcc tccaaagact tcaaaagatg agtgtaaaga
     5461 gtagatccaa gaccagttct ttgatgccta ggagacacgt aaacagtaga ctcagcagtc
     5521 caatcgtaag cattcctagc cttccaagga ccagcataag caataccagc cacttcacca
     5581 tccacttcag caacaagcca aggatacctc tctctaagcc tcacaagatc atcagtccac
     5641 tcttgtggtt cttgaggctc agtcctgaaa ttcacagtag aagtctcaat gtaatgattc
     5701 acaatagtgc acacagcagg catatcagcc tcagtagccc ttctaatatc agcaggcctt
     5761 ctttcaggag acatgcgaaa cgatccagat ccggtgcaga ttatttggat tgagagtgaa
     5821 tatgagactc taattggata ccgaggggaa tttatggaac gtcagtggag cattttgaca
     5881 agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc
     5941 tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctgagtggc tccttcaacg
     6001 ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat
     6061 aacgtgactc ccttaattct ccgctccgga tccgaattcg gcacagagtg ttcaacccca
     6121 gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc
     6181 acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc
     6241 accactcgat acaggcagcc catcaaccat gactactgtc cggtccaaca gatccgagca
     6301 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6361 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6421 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6481 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6541 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6601 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6661 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6721 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6781 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6841 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6901 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6961 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7021 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7081 ggtctgacag ctaaaacaat tcatccagta aaatataata ttttattttc tcccaatcag
     7141 gcttgatccc cagtaagtca aaaaatagct cgacatactg ttcttccccg atatcctccc
     7201 tgagcgaccg gacgcagaag gcaatgtcat accacttgtc cgccctgccg cttctcccaa
     7261 gatcaataaa gccacttact ttgccatctt tcacaaagat gttgctgtct cccaggtcgc
     7321 cgtgggaaaa gacaagttcc tcttcgggct tttccgtctt taaaaaatca tacagctcgc
     7381 gcggatcttt aaatggagtg tcctcttccc agttttcgca atccacatcg gccagatcgt
     7441 tattcagtaa gtaatccaat tcggctaagc ggctgtctaa gctattcgta tagggacaat
     7501 ccgatatgtc gatggagtga aagagcctga tgcactccgc atacagctcg ataatctttt
     7561 cagggctttg ttcatcttca tactcttccg agcaaaggac gccatcggcc tcactcatga
     7621 gcagattgct ccagccatca tgccgttcaa agtgcaggac ctttggaaca ggcagctttc
     7681 cttccagcca tagcatcatg tccttttccc gttccacatc ataggtggtc cctttatacc
     7741 ggctgtccgt catttttaaa tataggtttt cattttctcc caccagctta tataccttag
     7801 caggagacat tccttccgta tcttttacgc agcggtattt ttcgatcagt tttttcaatt
     7861 ccggtgatat tctcatttta gccatactct tcctttttca atattattga agcatttatc
     7921 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7981 gggttccgcg cactcggagc cttgatgtgg gcgctttttg cagctgttcg gctgtgcgct
     8041 ggccagacag ttatgcacag gccaggcggg ttttaagagt tttaataagt tttaaagagt
     8101 tttaggcgga aaaatcgcct tttttctctt ttatatcagt cacttacatg tgtgaccggt
     8161 tcccaatgta cggctttggg ttcccaatgt acgggttccg gttcccaatg tacggctttg
     8221 ggttcccaat gtacgtgcta tccacaggaa agaagctttt tcgacctttt tcccctgcta
     8281 gggcaatttg ccctagcatc tgctccgtac attaggaacc ggcggatgct tcgccctcga
     8341 tcaggttgcg gtagcgcatg actaggatcg ggccagcctg ccccgcctcc tccttcaaat
     8401 cgtactccgg caggtcattt gacccgatca gcttgcgcac ggtgaaacag aacttcttga
     8461 actctccggc gctgccactg cgttcgtaga tcgtcttgaa caaccatctg gcttctgcct
     8521 tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga tcgatcaaaa
     8581 agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc tcgcggtaca
     8641 tccaatcagc tagctcgatc tcgatgtact ccggccgccc ggtttcgctc tttacgatct
     8701 tgtagcggct aatcaaggct tcaccctcgg ataccgtcac caggcggccg ttcttggcct
     8761 tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt tctaccaggt
     8821 cgtctttctg ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc gcaacgtgtg
     8881 gacggaacac gcggccgggc ttgtctccct tcccttcccg gtatcggttc atggattcgg
     8941 ttagatggga aaccgccatc agtaccaggt cgtaatccca cacactggcc atgccggccg
     9001 gccctgcgga aacctctacg tgcccgtctg gaagctcgta gcggatcacc tcgccagctc
     9061 gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat gatgctgcga ctatcgcggg
     9121 tgcccacgtc atagagcatc ggaacgaaaa aatctggttg ctcgtcgccc ttgggcggct
     9181 tcctaatcga cgacgcaccg gctgccggcg gttgccggga ttctttgcgg attcgatcag
     9241 cggccgcttg ccacgattca ccggggcgtg cttctgcctc gatgcgttgc cgctgggcgg
     9301 cctgcgcggc cttcaacttc tccaccaggt catcacccag cgccgcgccg atttgtaccg
     9361 ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc ttgggggttc cagtgccatt
     9421 gcagggccgg cagacaaccc agccgcttac gcctggccaa ccgcccgttc ctccacacat
     9481 gggg
//
