LOCUS       sequence-342792-        8599 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 8599)
  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 8599)
  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_21362
FEATURES             Location/Qualifiers
     source          1..8599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    652..676
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(743..1450)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2064..2409)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2101..2121)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3442..3694)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4294..4476)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4330..4508)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4671..4695
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4831..5419)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4911..4930)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5590..6384)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6385..6489)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6551..6745
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6811..7884)
                     /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(8313..8599,1..343))
                     /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 tggcccatat gtcgtaaggg cttggctgca ccggaatcag cacgaagtcg gctgccttga
       61 tcgcggacac agccaagtcc gccgcctggg gcgctccgtc gatcactacg aagtcgcgcc
      121 ggccgatggc cttcacgtcg cggtcaatcg tcgggcggtc gatgccgaca acggttagcg
      181 gttgatcttc ccgcacggcc gcccaatcgc gggcactgcc ctggggatcg gaatcgacta
      241 acagaacatc ggccccggcg agttgcaggg cgcgggctag atgggttgcg atggtcgtct
      301 tgcctgaccc gcctttctgg ttaagtacag cgataacctt catgcgttcc ccttgcgtat
      361 ttgtttattt actcatcgca tcatatacgc agcgaccgca tgacgcaagc tgttttactc
      421 aaatacacat caccttttta gacggcggcg ctcggtttct tcagcggcca agctggccgg
      481 ccaggccgcc agcttggcat cagacaaacc ggccaggatt tcatgcagcc gcacggctga
      541 ctcccgaggg gaaccctgtg gttggcatgc acatacaaat ggacgaacgg ataaaccttt
      601 tcacgccctt ttaaatatcc gattattcta ataaacgctc ttttctctta ggtttacccg
      661 ccaatatatc ctgtcaaaca ctgatagttt aaacggatca cttcgtgtcc ctcggtcaca
      721 tgtgcatcct ctagtagcgg tcctgctgag cctcgacatg ttgtcgcaaa attcgccctg
      781 gacccgccca acgatttgtc gtcactgtca aggtttgacc tgcacttcat ttggggccca
      841 catacaccaa aaaaatgctg cataattctc ggggcagcaa gtcggttacc cggccgccgt
      901 gctggaccgg gttgaatggt gcccgtaact ttcggtagag cggacggcca atactcaact
      961 tcaaggaatc tcacccatgc gcgccggcgg ggaaccggag ttcccttcag tgagcgttat
     1021 tagttcgccg ctcggtgtgt cgtagatact agcccctggg gcacttttga aatttgaata
     1081 agatttatgt aatcagtctt ttaggtttga ccggttctgc cgcttttttt aaaattggat
     1141 ttgtaataat aaaacgcaat tgtttgttat tgtggcgctc tatcatagat gtcgctataa
     1201 acctattcag cacaatatat tgttttcatt ttaatattgt acatataagt agtagggtac
     1261 aatcagtaaa ttgaacggag aatattattc ataaaaatac gatagtaacg ggtgatatat
     1321 tcattagaat gaaccgaaac cggcggtaag gatctgagct acacatgctc aggtttttta
     1381 caacgtgcac aacagaattg aaagcaaata tcatgcgatc ataggcttct cgcatatctc
     1441 attaaagcag gacaagctca gatcaacaat gtcaaagcgt catactcctt agatgtctta
     1501 agagttcccc tagtgattcc aagcttgttt ttaagagtca cctgagtaag agtgtcgtcc
     1561 atgaactgac cgaagtactt ctgagccaaa gggttctcgt tgatctttgc caacattgtc
     1621 tcacctgtgt tagttgtagt agggttcttc accttcatca atgtgtcaac gtacattcta
     1681 gtctgcaaca aagttgagaa agcctcgtac tccttagagt ctggtgcagt gtccttagtg
     1741 atcctaagct tagacttcaa cacgtccaca gtccacttct cgtcgatgta ctgctcgaag
     1801 ttcctctgag ccaactggtt ctttcccaat ttagacataa ggttctgctc caaagtcata
     1861 gcgttctgtg gcttagcagc agtcactgcc tcgtccacct tcttcaaagc tccagtgttc
     1921 ttagtgaaag atgccaactt agaaagttgc ttagtcaaaa tctccatcct cattgtatcg
     1981 ataattgtaa atgtaattgt aatgttgttt gttgtttgtt gttgttggta attgttgtaa
     2041 aaatgagctc ttatactcga gcgtgtcctc tccaaatgaa atgaacttcc ttatatagag
     2101 gaagggtctt gcgaaggata gtgggattgt gcgtcatccc ttacgtcagt ggagatgtca
     2161 catcaatcca cttgctttgt agacgtggtt ggaacctctt ctttttccac gatgctcctc
     2221 gtgggtgggg gtccatcttt gggaccactg tcggcagaga gatcttgaat gatagccttt
     2281 cctttatcgc aatgatggca tttgtaggag ccaccttcct tttctactgt cctttcgatg
     2341 aagtgacaga tagctgggca atggaatccg aggaggtttc ccgaaattat cctttgttga
     2401 aaagtctcaa tagccctttg atcttctgag actgtatctt tgacattttt ggagtagacc
     2461 agagtgtcgt gctccaccat gttgacgaag attttcttct tgtcattgag tcgtaaaaga
     2521 ctctgtatga actgttcgcc agtcttgacg gcgagttctg ttagatcctc gatttgaatc
     2581 ttagactcca tgcatggcct tagattcagt aggaactacc tttttagaga ctccaatctc
     2641 tattacttgc cttggtttat gaagcaagcc ttgaatcgtc catactggaa tagtacttct
     2701 gatcttgaga aatatgtctt tctctgtgtt cttgatgcaa ttagtcctga atcttttgac
     2761 tgcatcttta accttcttgg gaaggtattt gatctcctgg agattgttac tcgggtagat
     2821 cgtcttgatg agacttgctg cgtaggcctc tctaaccatc tgtgggtcag cattctttct
     2881 gaaattgaag aggctaacct tctcattatc agtggtgaac atagtgtcgt caccttcacc
     2941 ttcgaacttc cttcctagat cgtaaagata gaggaaatcg tccattgtaa tctccggggc
     3001 aaaggagatc tcttttgggg ctggatcact gctgggcctt ttggttccta gcgtgagcca
     3061 gtgggctttt tgctttggtg ggcttgttag ggccttagca aagctcttgg gcttgagttg
     3121 agcttctcct ttggggatga agttcaacct gtctgtttgc tgacttgttg tgtacgcgtc
     3181 agctgctgct cttgcctctg taatagtggc aaatttcttg tgtgcaactc cgggaacgcc
     3241 gtttgttgcc gcctttgtac aaccccagtc atcgtatata ccggcatgtg gaccgttata
     3301 cacaacgtag tagttgatat gagggtgttg aatacccgat tctgctctga gaggagcaac
     3361 tgtgctgtta agctcagatt tttgtgggat tggaattcct ccaagcttga attcttgcag
     3421 cgctccaatt cagcaagcgt cgatctagta acatagatga caccgcgcgc gataatttat
     3481 cctagtttgc gcgctatatt ttgttttcta tcgcgtatta aatgtataat tgcgggactc
     3541 taatcataaa aacccatctc ataaataacg tcatgcatta catgttaatt attacatgct
     3601 taacgtaatt caacagaaat tatatgataa tcatcgcaag accggcaaca ggattcaatc
     3661 ttaagaaact ttattgccaa atgtttgaac gatctgcttg actctagatc cagagtcccg
     3721 ctcaaatctc agtcacagga agaacaggtc taggaggaac aggaagagag aaatcaagct
     3781 gccagaatcc cacatcatgc caatttccat gtttgaaacc agcagctcta agcattcccc
     3841 taggagcata accaagagcc tcatgcatcc tcacagaagg atcattagga agtccaatca
     3901 cagcaaccac agacttgaaa ccctgagcct ccaaagactt caaaagatga gtgtaaagag
     3961 tagatccaag accagttctt tgatgcctag gagacacgta aacagtagac tcagcagtcc
     4021 aatcgtaagc attcctagcc ttccaaggac cagcataagc aataccagcc acttcaccat
     4081 ccacttcagc aacaagccaa ggatacctct ctctaagcct cacaagatca tcagtccact
     4141 cttgtggttc ttgaggctca gtcctgaaat tcacagtaga agtctcaatg taatgattca
     4201 caatagtgca cacagcaggc atatcagcct cagtagccct tctaatatca gcaggccttc
     4261 tttcaggaga catgcgaaac gatccagatc cggtgcagat tatttggatt gagagtgaat
     4321 atgagactct aattggatac cgaggggaat ttatggaacg tcagtggagc attttgacaa
     4381 gaaatatttg ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct
     4441 gacgtatgtg cttagctcat taaactccag aaacccgcgg ctgagtggct ccttcaacgt
     4501 tgcggttctg tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata
     4561 acgtgactcc cttaattctc cgctccggat ccgaattcgg cacagagtgt tcaaccccag
     4621 tacattaaaa acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt gtttacacca
     4681 caatatatcc tgccaccagc cagccaacag ctccccgacc ggcagctcgg cacaaaatca
     4741 ccactcgata caggcagccc atcaaccatg actactgtcc ggtccaacag atccgagcag
     4801 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4861 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4921 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4981 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     5041 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     5101 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     5161 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     5221 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     5281 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     5341 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     5401 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     5461 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     5521 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     5581 gtctgacagc taaaacaatt catccagtaa aatataatat tttattttct cccaatcagg
     5641 cttgatcccc agtaagtcaa aaaatagctc gacatactgt tcttccccga tatcctccct
     5701 gagcgaccgg acgcagaagg caatgtcata ccacttgtcc gccctgccgc ttctcccaag
     5761 atcaataaag ccacttactt tgccatcttt cacaaagatg ttgctgtctc ccaggtcgcc
     5821 gtgggaaaag acaagttcct cttcgggctt ttccgtcttt aaaaaatcat acagctcgcg
     5881 cggatcttta aatggagtgt cctcttccca gttttcgcaa tccacatcgg ccagatcgtt
     5941 attcagtaag taatccaatt cggctaagcg gctgtctaag ctattcgtat agggacaatc
     6001 cgatatgtcg atggagtgaa agagcctgat gcactccgca tacagctcga taatcttttc
     6061 agggctttgt tcatcttcat actcttccga gcaaaggacg ccatcggcct cactcatgag
     6121 cagattgctc cagccatcat gccgttcaaa gtgcaggacc tttggaacag gcagctttcc
     6181 ttccagccat agcatcatgt ccttttcccg ttccacatca taggtggtcc ctttataccg
     6241 gctgtccgtc atttttaaat ataggttttc attttctccc accagcttat ataccttagc
     6301 aggagacatt ccttccgtat cttttacgca gcggtatttt tcgatcagtt ttttcaattc
     6361 cggtgatatt ctcattttag ccatactctt cctttttcaa tattattgaa gcatttatca
     6421 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6481 ggttccgcgc actcggagcc ttgatgtggg cgctttttgc agctgttcgg ctgtgcgctg
     6541 gccagacagt tatgcacagg ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt
     6601 ttaggcggaa aaatcgcctt ttttctcttt tatatcagtc acttacatgt gtgaccggtt
     6661 cccaatgtac ggctttgggt tcccaatgta cgggttccgg ttcccaatgt acggctttgg
     6721 gttcccaatg tacgtgctat ccacaggaaa gaagcttttt cgaccttttt cccctgctag
     6781 ggcaatttgc cctagcatct gctccgtaca ttaggaaccg gcggatgctt cgccctcgat
     6841 caggttgcgg tagcgcatga ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc
     6901 gtactccggc aggtcatttg acccgatcag cttgcgcacg gtgaaacaga acttcttgaa
     6961 ctctccggcg ctgccactgc gttcgtagat cgtcttgaac aaccatctgg cttctgcctt
     7021 gcctgcggcg cggcgtgcca ggcggtagag aaaacggccg atgccgggat cgatcaaaaa
     7081 gtaatcgggg tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct cgcggtacat
     7141 ccaatcagct agctcgatct cgatgtactc cggccgcccg gtttcgctct ttacgatctt
     7201 gtagcggcta atcaaggctt caccctcgga taccgtcacc aggcggccgt tcttggcctt
     7261 cttcgtacgc tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc
     7321 gtctttctgc tttccgccat cggctcgccg gcagaacttg agtacgtccg caacgtgtgg
     7381 acggaacacg cggccgggct tgtctccctt cccttcccgg tatcggttca tggattcggt
     7441 tagatgggaa accgccatca gtaccaggtc gtaatcccac acactggcca tgccggccgg
     7501 ccctgcggaa acctctacgt gcccgtctgg aagctcgtag cggatcacct cgccagctcg
     7561 tcggtcacgc ttcgacagac ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt
     7621 gcccacgtca tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt
     7681 cctaatcgac gacgcaccgg ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc
     7741 ggccgcttgc cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc
     7801 ctgcgcggcc ttcaacttct ccaccaggtc atcacccagc gccgcgccga tttgtaccgg
     7861 gccggatggt ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc agtgccattg
     7921 cagggccggc agacaaccca gccgcttacg cctggccaac cgcccgttcc tccacacatg
     7981 gggcattcca cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc tcctttagcc
     8041 gctaaaattc atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgta
     8101 ttcagatagc agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt
     8161 gtgatcctcc gccggcaact gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct
     8221 ggccaacgtt gcagccttgc tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt
     8281 gcttttgctc attttctctt tacctcatta actcaaatga gttttgattt aatttcagcg
     8341 gccagcgcct ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg
     8401 ccggcggcgg cagtgcctgg gtagctcacg cgctgcgtga tacgggactc aagaatgggc
     8461 agctcgtacc cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc
     8521 cgcgacacga caaaggccgc ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc
     8581 agctccacca ggtcggcgg
//
