LOCUS       sequence-342804-        9079 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 9079)
  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 9079)
  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_11947
FEATURES             Location/Qualifiers
     source          1..9079
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(145..327)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(181..359)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    522..546
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(682..1270)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(762..781)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1441..2235)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(2236..2340)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2402..2596
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2662..3735)
                     /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(4164..4793)
                     /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    5102..5126
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(5193..5900)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(6994..7339)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(7031..7051)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(8372..8624)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 cttgaggctc agtcctgaaa ttcacagtag aagtctcaat gtaatgattc acaatagtgc
       61 acacagcagg catatcagcc tcagtagccc ttctaatatc agcaggcctt ctttcaggag
      121 acatgcgaaa cgatccagat ccggtgcaga ttatttggat tgagagtgaa tatgagactc
      181 taattggata ccgaggggaa tttatggaac gtcagtggag cattttgaca agaaatattt
      241 gctagctgat agtgacctta ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt
      301 gcttagctca ttaaactcca gaaacccgcg gctgagtggc tccttcaacg ttgcggttct
      361 gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc
      421 ccttaattct ccgctccgga tccgaattcg gcacagagtg ttcaacccca gtacattaaa
      481 aacgtccgca atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc
      541 ctgccaccag ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat
      601 acaggcagcc catcaaccat gactactgtc cggtccaaca gatccgagca ggaaccgtaa
      661 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      721 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      781 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      841 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      901 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      961 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1021 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1081 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1141 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1201 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1261 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1321 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1381 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1441 ctaaaacaat tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc
     1501 cagtaagtca aaaaatagct cgacatactg ttcttccccg atatcctccc tgagcgaccg
     1561 gacgcagaag gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa
     1621 gccacttact ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa
     1681 gacaagttcc tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt
     1741 aaatggagtg tcctcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa
     1801 gtaatccaat tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc
     1861 gatggagtga aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg
     1921 ttcatcttca tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct
     1981 ccagccatca tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca
     2041 tagcatcatg tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt
     2101 catttttaaa tataggtttt cattttctcc caccagctta tataccttag caggagacat
     2161 tccttccgta tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat
     2221 tctcatttta gccatactct tcctttttca atattattga agcatttatc agggttattg
     2281 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     2341 cactcggagc cttgatgtgg gcgctttttg cagctgttcg gctgtgcgct ggccagacag
     2401 ttatgcacag gccaggcggg ttttaagagt tttaataagt tttaaagagt tttaggcgga
     2461 aaaatcgcct tttttctctt ttatatcagt cacttacatg tgtgaccggt tcccaatgta
     2521 cggctttggg ttcccaatgt acgggttccg gttcccaatg tacggctttg ggttcccaat
     2581 gtacgtgcta tccacaggaa agaagctttt tcgacctttt tcccctgcta gggcaatttg
     2641 ccctagcatc tgctccgtac attaggaacc ggcggatgct tcgccctcga tcaggttgcg
     2701 gtagcgcatg actaggatcg ggccagcctg ccccgcctcc tccttcaaat cgtactccgg
     2761 caggtcattt gacccgatca gcttgcgcac ggtgaaacag aacttcttga actctccggc
     2821 gctgccactg cgttcgtaga tcgtcttgaa caaccatctg gcttctgcct tgcctgcggc
     2881 gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga tcgatcaaaa agtaatcggg
     2941 gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc tcgcggtaca tccaatcagc
     3001 tagctcgatc tcgatgtact ccggccgccc ggtttcgctc tttacgatct tgtagcggct
     3061 aatcaaggct tcaccctcgg ataccgtcac caggcggccg ttcttggcct tcttcgtacg
     3121 ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt tctaccaggt cgtctttctg
     3181 ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc gcaacgtgtg gacggaacac
     3241 gcggccgggc ttgtctccct tcccttcccg gtatcggttc atggattcgg ttagatggga
     3301 aaccgccatc agtaccaggt cgtaatccca cacactggcc atgccggccg gccctgcgga
     3361 aacctctacg tgcccgtctg gaagctcgta gcggatcacc tcgccagctc gtcggtcacg
     3421 cttcgacaga cggaaaacgg ccacgtccat gatgctgcga ctatcgcggg tgcccacgtc
     3481 atagagcatc ggaacgaaaa aatctggttg ctcgtcgccc ttgggcggct tcctaatcga
     3541 cgacgcaccg gctgccggcg gttgccggga ttctttgcgg attcgatcag cggccgcttg
     3601 ccacgattca ccggggcgtg cttctgcctc gatgcgttgc cgctgggcgg cctgcgcggc
     3661 cttcaacttc tccaccaggt catcacccag cgccgcgccg atttgtaccg ggccggatgg
     3721 tttgcgaccg ctcacgccga ttcctcgggc ttgggggttc cagtgccatt gcagggccgg
     3781 cagacaaccc agccgcttac gcctggccaa ccgcccgttc ctccacacat ggggcattcc
     3841 acggcgtcgg tgcctggttg ttcttgattt tccatgccgc ctcctttagc cgctaaaatt
     3901 catctactca tttattcatt tgctcattta ctctggtagc tgcgcgatgt attcagatag
     3961 cagctcggta atggtcttgc cttggcgtac cgcgtacatc ttcagcttgg tgtgatcctc
     4021 cgccggcaac tgaaagttga cccgcttcat ggctggcgtg tctgccaggc tggccaacgt
     4081 tgcagccttg ctgctgcgtg cgctcggacg gccggcactt agcgtgtttg tgcttttgct
     4141 cattttctct ttacctcatt aactcaaatg agttttgatt taatttcagc ggccagcgcc
     4201 tggacctcgc gggcagcgtc gccctcgggt tctgattcaa gaacggttgt gccggcggcg
     4261 gcagtgcctg ggtagctcac gcgctgcgtg atacgggact caagaatggg cagctcgtac
     4321 ccggccagcg cctcggcaac ctcaccgccg atgcgcgtgc ctttgatcgc ccgcgacacg
     4381 acaaaggccg cttgtagcct tccatccgtg acctcaatgc gctgcttaac cagctccacc
     4441 aggtcggcgg tggcccatat gtcgtaaggg cttggctgca ccggaatcag cacgaagtcg
     4501 gctgccttga tcgcggacac agccaagtcc gccgcctggg gcgctccgtc gatcactacg
     4561 aagtcgcgcc ggccgatggc cttcacgtcg cggtcaatcg tcgggcggtc gatgccgaca
     4621 acggttagcg gttgatcttc ccgcacggcc gcccaatcgc gggcactgcc ctggggatcg
     4681 gaatcgacta acagaacatc ggccccggcg agttgcaggg cgcgggctag atgggttgcg
     4741 atggtcgtct tgcctgaccc gcctttctgg ttaagtacag cgataacctt catgcgttcc
     4801 ccttgcgtat ttgtttattt actcatcgca tcatatacgc agcgaccgca tgacgcaagc
     4861 tgttttactc aaatacacat caccttttta gacggcggcg ctcggtttct tcagcggcca
     4921 agctggccgg ccaggccgcc agcttggcat cagacaaacc ggccaggatt tcatgcagcc
     4981 gcacggctga ctcccgaggg gaaccctgtg gttggcatgc acatacaaat ggacgaacgg
     5041 ataaaccttt tcacgccctt ttaaatatcc gattattcta ataaacgctc ttttctctta
     5101 ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt aaacggatca cttcgtgtcc
     5161 ctcggtcaca tgtgcatcct ctagtagcgg tcctgctgag cctcgacatg ttgtcgcaaa
     5221 attcgccctg gacccgccca acgatttgtc gtcactgtca aggtttgacc tgcacttcat
     5281 ttggggccca catacaccaa aaaaatgctg cataattctc ggggcagcaa gtcggttacc
     5341 cggccgccgt gctggaccgg gttgaatggt gcccgtaact ttcggtagag cggacggcca
     5401 atactcaact tcaaggaatc tcacccatgc gcgccggcgg ggaaccggag ttcccttcag
     5461 tgagcgttat tagttcgccg ctcggtgtgt cgtagatact agcccctggg gcacttttga
     5521 aatttgaata agatttatgt aatcagtctt ttaggtttga ccggttctgc cgcttttttt
     5581 aaaattggat ttgtaataat aaaacgcaat tgtttgttat tgtggcgctc tatcatagat
     5641 gtcgctataa acctattcag cacaatatat tgttttcatt ttaatattgt acatataagt
     5701 agtagggtac aatcagtaaa ttgaacggag aatattattc ataaaaatac gatagtaacg
     5761 ggtgatatat tcattagaat gaaccgaaac cggcggtaag gatctgagct acacatgctc
     5821 aggtttttta caacgtgcac aacagaattg aaagcaaata tcatgcgatc ataggcttct
     5881 cgcatatctc attaaagcag gacaagctca agtagatccc tcagtgtttc caccgttgtc
     5941 acctcctgtg ttacctccct cgtcaccacc aatgtctcca ccagtatcac cacctttgtc
     6001 tcctcctgta tttccaccat tgtctcctcc attatcaggc tctgccatct caagtgatcc
     6061 gcactcagag tcatcctcag ccatgtccaa tgatccacat tctgagtcat cctctgccat
     6121 ctccaatgaa ccacattcag aatctccctc tccaccaatc tcagcttctc caccgatttc
     6181 tccctgctga acagagtcac cagtgttttc acctcctgac tcttttttat ctcctccgtt
     6241 gtcacctcca gattctttat tatcaccacc agatttctta ttatcacctc cattgtcacc
     6301 accttgctgc ttattgtcac cacctttgtc acctcctgag tcgatgttat caccaccaac
     6361 ctccttgttg tcacctccca cctctttgtt gtcacctcca gactccttct tatctccacc
     6421 tgattcctta ttatctcctc cattatcacc tcctccgtta tctccagaag attgcttgtt
     6481 atctcctcca ttatcacctc ctgattcttt attgtcacca ccagactcct tgttatctcc
     6541 acctgactct ttgttgtctc ctgtattaga acctcctccc cagttgtttg tacctccacc
     6601 agagttccac tggttgtttg aagaaccacc agtatttgta gatggagcct gtgtccaagt
     6661 attaccacct gtatctcctc ccttgtctcc tccagtatca gttgaaggag cctgagtcca
     6721 tgtgtttcca ccagtgtctc ctcccttgtc accacctgta tctcctccga tgtctccacc
     6781 gttatcaggc tcagccatct caagtgatcc acactcagag tcgtcctctg ccatctcaag
     6841 agaaccgcac tcagaatcgt cctctgccat ttccaaagaa ccacactctg agtcatcagc
     6901 cattgtatcg ataattgtaa atgtaattgt aatgttgttt gttgtttgtt gttgttggta
     6961 attgttgtaa aaatgagctc ttatactcga gcgtgtcctc tccaaatgaa atgaacttcc
     7021 ttatatagag gaagggtctt gcgaaggata gtgggattgt gcgtcatccc ttacgtcagt
     7081 ggagatgtca catcaatcca cttgctttgt agacgtggtt ggaacctctt ctttttccac
     7141 gatgctcctc gtgggtgggg gtccatcttt gggaccactg tcggcagaga gatcttgaat
     7201 gatagccttt cctttatcgc aatgatggca tttgtaggag ccaccttcct tttctactgt
     7261 cctttcgatg aagtgacaga tagctgggca atggaatccg aggaggtttc ccgaaattat
     7321 cctttgttga aaagtctcaa tagccctttg atcttctgag actgtatctt tgacattttt
     7381 ggagtagacc agagtgtcgt gctccaccat gttgacgaag attttcttct tgtcattgag
     7441 tcgtaaaaga ctctgtatga actgttcgcc agtcttgacg gcgagttctg ttagatcctc
     7501 gatttgaatc ttagactcca tgcatggcct tagattcagt aggaactacc tttttagaga
     7561 ctccaatctc tattacttgc cttggtttat gaagcaagcc ttgaatcgtc catactggaa
     7621 tagtacttct gatcttgaga aatatgtctt tctctgtgtt cttgatgcaa ttagtcctga
     7681 atcttttgac tgcatcttta accttcttgg gaaggtattt gatctcctgg agattgttac
     7741 tcgggtagat cgtcttgatg agacttgctg cgtaggcctc tctaaccatc tgtgggtcag
     7801 cattctttct gaaattgaag aggctaacct tctcattatc agtggtgaac atagtgtcgt
     7861 caccttcacc ttcgaacttc cttcctagat cgtaaagata gaggaaatcg tccattgtaa
     7921 tctccggggc aaaggagatc tcttttgggg ctggatcact gctgggcctt ttggttccta
     7981 gcgtgagcca gtgggctttt tgctttggtg ggcttgttag ggccttagca aagctcttgg
     8041 gcttgagttg agcttctcct ttggggatga agttcaacct gtctgtttgc tgacttgttg
     8101 tgtacgcgtc agctgctgct cttgcctctg taatagtggc aaatttcttg tgtgcaactc
     8161 cgggaacgcc gtttgttgcc gcctttgtac aaccccagtc atcgtatata ccggcatgtg
     8221 gaccgttata cacaacgtag tagttgatat gagggtgttg aatacccgat tctgctctga
     8281 gaggagcaac tgtgctgtta agctcagatt tttgtgggat tggaattcct ccaagcttga
     8341 attcttgcag cgctccaatt cagcaagcgt cgatctagta acatagatga caccgcgcgc
     8401 gataatttat cctagtttgc gcgctatatt ttgttttcta tcgcgtatta aatgtataat
     8461 tgcgggactc taatcataaa aacccatctc ataaataacg tcatgcatta catgttaatt
     8521 attacatgct taacgtaatt caacagaaat tatatgataa tcatcgcaag accggcaaca
     8581 ggattcaatc ttaagaaact ttattgccaa atgtttgaac gatctgcttg actctagatc
     8641 cagagtcccg ctcaaatctc agtcacagga agaacaggtc taggaggaac aggaagagag
     8701 aaatcaagct gccagaatcc cacatcatgc caatttccat gtttgaaacc agcagctcta
     8761 agcattcccc taggagcata accaagagcc tcatgcatcc tcacagaagg atcattagga
     8821 agtccaatca cagcaaccac agacttgaaa ccctgagcct ccaaagactt caaaagatga
     8881 gtgtaaagag tagatccaag accagttctt tgatgcctag gagacacgta aacagtagac
     8941 tcagcagtcc aatcgtaagc attcctagcc ttccaaggac cagcataagc aataccagcc
     9001 acttcaccat ccacttcagc aacaagccaa ggatacctct ctctaagcct cacaagatca
     9061 tcagtccact cttgtggtt
//
