LOCUS       sequence-342753-        9940 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 9940)
  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 9940)
  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_16705
FEATURES             Location/Qualifiers
     source          1..9940
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    632..656
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(723..1430)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3385..3730)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3422..3442)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4763..5015)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5615..5797)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5651..5829)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5992..6016
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6152..6740)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6232..6251)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6911..7705)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7706..7810)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7872..8066
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8132..9205)
                     /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(9634..9940,1..323))
                     /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 cttggctgca ccggaatcag cacgaagtcg gctgccttga tcgcggacac agccaagtcc
       61 gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg
      121 cggtcaatcg tcgggcggtc gatgccgaca acggttagcg gttgatcttc ccgcacggcc
      181 gcccaatcgc gggcactgcc ctggggatcg gaatcgacta acagaacatc ggccccggcg
      241 agttgcaggg cgcgggctag atgggttgcg atggtcgtct tgcctgaccc gcctttctgg
      301 ttaagtacag cgataacctt catgcgttcc ccttgcgtat ttgtttattt actcatcgca
      361 tcatatacgc agcgaccgca tgacgcaagc tgttttactc aaatacacat caccttttta
      421 gacggcggcg ctcggtttct tcagcggcca agctggccgg ccaggccgcc agcttggcat
      481 cagacaaacc ggccaggatt tcatgcagcc gcacggctga ctcccgaggg gaaccctgtg
      541 gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc
      601 gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca
      661 ctgatagttt aaacggatca cttcgtgtcc ctcggtcaca tgtgcatcct ctagtagcgg
      721 tcctgctgag cctcgacatg ttgtcgcaaa attcgccctg gacccgccca acgatttgtc
      781 gtcactgtca aggtttgacc tgcacttcat ttggggccca catacaccaa aaaaatgctg
      841 cataattctc ggggcagcaa gtcggttacc cggccgccgt gctggaccgg gttgaatggt
      901 gcccgtaact ttcggtagag cggacggcca atactcaact tcaaggaatc tcacccatgc
      961 gcgccggcgg ggaaccggag ttcccttcag tgagcgttat tagttcgccg ctcggtgtgt
     1021 cgtagatact agcccctggg gcacttttga aatttgaata agatttatgt aatcagtctt
     1081 ttaggtttga ccggttctgc cgcttttttt aaaattggat ttgtaataat aaaacgcaat
     1141 tgtttgttat tgtggcgctc tatcatagat gtcgctataa acctattcag cacaatatat
     1201 tgttttcatt ttaatattgt acatataagt agtagggtac aatcagtaaa ttgaacggag
     1261 aatattattc ataaaaatac gatagtaacg ggtgatatat tcattagaat gaaccgaaac
     1321 cggcggtaag gatctgagct acacatgctc aggtttttta caacgtgcac aacagaattg
     1381 aaagcaaata tcatgcgatc ataggcttct cgcatatctc attaaagcag gacaagctca
     1441 aggagtagat gatgccactg ccaacaactt gatgtacctg tccaacattt cgtctgctgc
     1501 aggcacacca acgaacaccc tcttaagctc ctcaggagtt ttctttgcgt cttgccactt
     1561 caagataagt gcagactgca agtcagtagc aagcttctct gttgaagggt cgttctttgc
     1621 agcagagaac atcctcaaca atgcaatatc gtgtgaatta tccctaagtc cgtcgataac
     1681 agtagttttt tcgtttgggt acctgtcatt gaaagtttta aggtattgca cccaagtgtt
     1741 aaatttaggg tcagccaaaa gcttctcacc agactcatcc aagtgcaaag acctgaatgc
     1801 agtcttaggc tccatcttga tgtccaacca atagttgtgg tatgcgttct caaccttctc
     1861 agccatcaac ctagtaagag ggttcttcct acctgtctcg atgatcctct ccactccgaa
     1921 aggctggtat gagatcctca acatgtcaaa ccaagactct tggttattag ggtgttcttt
     1981 attaaacttt tgaaggtatg ccaaccactt atgaaacaaa gtattagaca agatatcgtc
     2041 accaacgttg tcaagagcca acaccttgaa gataccctca ggagactttc ctgaagcgaa
     2101 aagagtagat gccttctcca tttgcaattt tgtagctgta gattcaaggt ttggaaattt
     2161 cttagcctct cccaacaccc taaacaacct catgtctccg aatgtgtcac ccaactccaa
     2221 gaacatttta tccacctttg atgggttctt atcaacgaac acattcatgt atgaaaccca
     2281 cacgttcaac tgaggctcgt tgaagaaagc tgtatcaccg tatgatgatc taagcctgtt
     2341 gtacacatca agtggagact ccttaagatt catccacctg tcgaactgct gagcctgcaa
     2401 cttagcagcc aaagtcttag tctctccctt agatgatgct ccgataagag tgtccaacaa
     2461 ctgagcatca ccgtacttcc tagtaagtgt ctcgattgca gtagccttct tctctgggta
     2521 cctagcattg tatgcagaaa ggtacctctc ccactccttc caccttggag aaccaaggaa
     2581 tgcttgtgta gttgttccat atttgaatgc gtccatcacc ttgtctggag tctctcttga
     2641 gttcaaccag aagtcgaaaa gagctgactc caccttatta gccacagtct tagcctctgt
     2701 agtagcctta ccgtttgcaa taagcttaag caacacgtca gttcctgcct cgtcacctct
     2761 tgagaacctc tgcttaagtg tagggatgat agacaaaggt gcctcctttg ggtgccttgc
     2821 attcacatca tccacgtact taagccaagc tgaaaattca ggcttaggga agatagagat
     2881 tccagcccag ttaaggttaa gagcctcgaa caccactgaa ggatccttcc ttgtagcgat
     2941 ccagaacctg atctggtctt tctccaattt aagtgccaat tcagaaagtt gagggtacaa
     3001 tttagctgcc tggatagtgt cgtaaaccac cttgtctccg tgaagtgaag tcaatgttga
     3061 gatcactgaa gattctttgt gagatgattt tgcagacaaa tcatcagcgt actgaaccca
     3121 ctttgcgaaa ttaggctcgt gcaacaatga aaaaatatta ggctcgtcaa ggttcatatt
     3181 tttgaacaca agaccagcag gaagcctctc atcaagccat tgctgaagct ttgctggagt
     3241 cacagctgat gacttcacca aagccttaat attttccacg attgaaggag ccattgtatc
     3301 gataattgta aatgtaattg taatgttgtt tgttgtttgt tgttgttggt aattgttgta
     3361 aaaatgagct cttatactcg agcgtgtcct ctccaaatga aatgaacttc cttatataga
     3421 ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatgtc
     3481 acatcaatcc acttgctttg tagacgtggt tggaacctct tctttttcca cgatgctcct
     3541 cgtgggtggg ggtccatctt tgggaccact gtcggcagag agatcttgaa tgatagcctt
     3601 tcctttatcg caatgatggc atttgtagga gccaccttcc ttttctactg tcctttcgat
     3661 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgaaatta tcctttgttg
     3721 aaaagtctca atagcccttt gatcttctga gactgtatct ttgacatttt tggagtagac
     3781 cagagtgtcg tgctccacca tgttgacgaa gattttcttc ttgtcattga gtcgtaaaag
     3841 actctgtatg aactgttcgc cagtcttgac ggcgagttct gttagatcct cgatttgaat
     3901 cttagactcc atgcatggcc ttagattcag taggaactac ctttttagag actccaatct
     3961 ctattacttg ccttggttta tgaagcaagc cttgaatcgt ccatactgga atagtacttc
     4021 tgatcttgag aaatatgtct ttctctgtgt tcttgatgca attagtcctg aatcttttga
     4081 ctgcatcttt aaccttcttg ggaaggtatt tgatctcctg gagattgtta ctcgggtaga
     4141 tcgtcttgat gagacttgct gcgtaggcct ctctaaccat ctgtgggtca gcattctttc
     4201 tgaaattgaa gaggctaacc ttctcattat cagtggtgaa catagtgtcg tcaccttcac
     4261 cttcgaactt ccttcctaga tcgtaaagat agaggaaatc gtccattgta atctccgggg
     4321 caaaggagat ctcttttggg gctggatcac tgctgggcct tttggttcct agcgtgagcc
     4381 agtgggcttt ttgctttggt gggcttgtta gggccttagc aaagctcttg ggcttgagtt
     4441 gagcttctcc tttggggatg aagttcaacc tgtctgtttg ctgacttgtt gtgtacgcgt
     4501 cagctgctgc tcttgcctct gtaatagtgg caaatttctt gtgtgcaact ccgggaacgc
     4561 cgtttgttgc cgcctttgta caaccccagt catcgtatat accggcatgt ggaccgttat
     4621 acacaacgta gtagttgata tgagggtgtt gaatacccga ttctgctctg agaggagcaa
     4681 ctgtgctgtt aagctcagat ttttgtggga ttggaattcc tccaagcttg aattcttgca
     4741 gcgctccaat tcagcaagcg tcgatctagt aacatagatg acaccgcgcg cgataattta
     4801 tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact
     4861 ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc
     4921 ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat
     4981 cttaagaaac tttattgcca aatgtttgaa cgatctgctt gactctagat ccagagtccc
     5041 gctcaaatct cagtcacagg aagaacaggt ctaggaggaa caggaagaga gaaatcaagc
     5101 tgccagaatc ccacatcatg ccaatttcca tgtttgaaac cagcagctct aagcattccc
     5161 ctaggagcat aaccaagagc ctcatgcatc ctcacagaag gatcattagg aagtccaatc
     5221 acagcaacca cagacttgaa accctgagcc tccaaagact tcaaaagatg agtgtaaaga
     5281 gtagatccaa gaccagttct ttgatgccta ggagacacgt aaacagtaga ctcagcagtc
     5341 caatcgtaag cattcctagc cttccaagga ccagcataag caataccagc cacttcacca
     5401 tccacttcag caacaagcca aggatacctc tctctaagcc tcacaagatc atcagtccac
     5461 tcttgtggtt cttgaggctc agtcctgaaa ttcacagtag aagtctcaat gtaatgattc
     5521 acaatagtgc acacagcagg catatcagcc tcagtagccc ttctaatatc agcaggcctt
     5581 ctttcaggag acatgcgaaa cgatccagat ccggtgcaga ttatttggat tgagagtgaa
     5641 tatgagactc taattggata ccgaggggaa tttatggaac gtcagtggag cattttgaca
     5701 agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc
     5761 tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctgagtggc tccttcaacg
     5821 ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat
     5881 aacgtgactc ccttaattct ccgctccgga tccgaattcg gcacagagtg ttcaacccca
     5941 gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc
     6001 acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc
     6061 accactcgat acaggcagcc catcaaccat gactactgtc cggtccaaca gatccgagca
     6121 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6181 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6241 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6301 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6361 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6421 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6481 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6541 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6601 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6661 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6721 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6781 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6841 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6901 ggtctgacag ctaaaacaat tcatccagta aaatataata ttttattttc tcccaatcag
     6961 gcttgatccc cagtaagtca aaaaatagct cgacatactg ttcttccccg atatcctccc
     7021 tgagcgaccg gacgcagaag gcaatgtcat accacttgtc cgccctgccg cttctcccaa
     7081 gatcaataaa gccacttact ttgccatctt tcacaaagat gttgctgtct cccaggtcgc
     7141 cgtgggaaaa gacaagttcc tcttcgggct tttccgtctt taaaaaatca tacagctcgc
     7201 gcggatcttt aaatggagtg tcctcttccc agttttcgca atccacatcg gccagatcgt
     7261 tattcagtaa gtaatccaat tcggctaagc ggctgtctaa gctattcgta tagggacaat
     7321 ccgatatgtc gatggagtga aagagcctga tgcactccgc atacagctcg ataatctttt
     7381 cagggctttg ttcatcttca tactcttccg agcaaaggac gccatcggcc tcactcatga
     7441 gcagattgct ccagccatca tgccgttcaa agtgcaggac ctttggaaca ggcagctttc
     7501 cttccagcca tagcatcatg tccttttccc gttccacatc ataggtggtc cctttatacc
     7561 ggctgtccgt catttttaaa tataggtttt cattttctcc caccagctta tataccttag
     7621 caggagacat tccttccgta tcttttacgc agcggtattt ttcgatcagt tttttcaatt
     7681 ccggtgatat tctcatttta gccatactct tcctttttca atattattga agcatttatc
     7741 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7801 gggttccgcg cactcggagc cttgatgtgg gcgctttttg cagctgttcg gctgtgcgct
     7861 ggccagacag ttatgcacag gccaggcggg ttttaagagt tttaataagt tttaaagagt
     7921 tttaggcgga aaaatcgcct tttttctctt ttatatcagt cacttacatg tgtgaccggt
     7981 tcccaatgta cggctttggg ttcccaatgt acgggttccg gttcccaatg tacggctttg
     8041 ggttcccaat gtacgtgcta tccacaggaa agaagctttt tcgacctttt tcccctgcta
     8101 gggcaatttg ccctagcatc tgctccgtac attaggaacc ggcggatgct tcgccctcga
     8161 tcaggttgcg gtagcgcatg actaggatcg ggccagcctg ccccgcctcc tccttcaaat
     8221 cgtactccgg caggtcattt gacccgatca gcttgcgcac ggtgaaacag aacttcttga
     8281 actctccggc gctgccactg cgttcgtaga tcgtcttgaa caaccatctg gcttctgcct
     8341 tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga tcgatcaaaa
     8401 agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc tcgcggtaca
     8461 tccaatcagc tagctcgatc tcgatgtact ccggccgccc ggtttcgctc tttacgatct
     8521 tgtagcggct aatcaaggct tcaccctcgg ataccgtcac caggcggccg ttcttggcct
     8581 tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt tctaccaggt
     8641 cgtctttctg ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc gcaacgtgtg
     8701 gacggaacac gcggccgggc ttgtctccct tcccttcccg gtatcggttc atggattcgg
     8761 ttagatggga aaccgccatc agtaccaggt cgtaatccca cacactggcc atgccggccg
     8821 gccctgcgga aacctctacg tgcccgtctg gaagctcgta gcggatcacc tcgccagctc
     8881 gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat gatgctgcga ctatcgcggg
     8941 tgcccacgtc atagagcatc ggaacgaaaa aatctggttg ctcgtcgccc ttgggcggct
     9001 tcctaatcga cgacgcaccg gctgccggcg gttgccggga ttctttgcgg attcgatcag
     9061 cggccgcttg ccacgattca ccggggcgtg cttctgcctc gatgcgttgc cgctgggcgg
     9121 cctgcgcggc cttcaacttc tccaccaggt catcacccag cgccgcgccg atttgtaccg
     9181 ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc ttgggggttc cagtgccatt
     9241 gcagggccgg cagacaaccc agccgcttac gcctggccaa ccgcccgttc ctccacacat
     9301 ggggcattcc acggcgtcgg tgcctggttg ttcttgattt tccatgccgc ctcctttagc
     9361 cgctaaaatt catctactca tttattcatt tgctcattta ctctggtagc tgcgcgatgt
     9421 attcagatag cagctcggta atggtcttgc cttggcgtac cgcgtacatc ttcagcttgg
     9481 tgtgatcctc cgccggcaac tgaaagttga cccgcttcat ggctggcgtg tctgccaggc
     9541 tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg gccggcactt agcgtgtttg
     9601 tgcttttgct cattttctct ttacctcatt aactcaaatg agttttgatt taatttcagc
     9661 ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt tctgattcaa gaacggttgt
     9721 gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg atacgggact caagaatggg
     9781 cagctcgtac ccggccagcg cctcggcaac ctcaccgccg atgcgcgtgc ctttgatcgc
     9841 ccgcgacacg acaaaggccg cttgtagcct tccatccgtg acctcaatgc gctgcttaac
     9901 cagctccacc aggtcggcgg tggcccatat gtcgtaaggg
//
