LOCUS       sequence-342334-        8692 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 8692)
  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 8692)
  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_13044
FEATURES             Location/Qualifiers
     source          1..8692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    754..778
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(845..1552)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2259..2604)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2296..2316)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3637..3889)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4489..4671)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4525..4703)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4866..4890
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5026..5614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5106..5125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5785..6579)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6580..6684)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6746..6940
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7006..8079)
                     /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(8508..8692,1..445))
                     /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 cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg
       61 tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag
      121 ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt
      181 ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt
      241 cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg
      301 ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg
      361 cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct
      421 ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg
      481 catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt
      541 tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc
      601 atcagacaaa ccggccagga tttcatgcag ccgcacggct gactcccgag gggaaccctg
      661 tggttggcat gcacatacaa atggacgaac ggataaacct tttcacgccc ttttaaatat
      721 ccgattattc taataaacgc tcttttctct taggtttacc cgccaatata tcctgtcaaa
      781 cactgatagt ttaaacggat cacttcgtgt ccctcggtca catgtgcatc ctctagtagc
      841 ggtcctgctg agcctcgaca tgttgtcgca aaattcgccc tggacccgcc caacgatttg
      901 tcgtcactgt caaggtttga cctgcacttc atttggggcc cacatacacc aaaaaaatgc
      961 tgcataattc tcggggcagc aagtcggtta cccggccgcc gtgctggacc gggttgaatg
     1021 gtgcccgtaa ctttcggtag agcggacggc caatactcaa cttcaaggaa tctcacccat
     1081 gcgcgccggc ggggaaccgg agttcccttc agtgagcgtt attagttcgc cgctcggtgt
     1141 gtcgtagata ctagcccctg gggcactttt gaaatttgaa taagatttat gtaatcagtc
     1201 ttttaggttt gaccggttct gccgcttttt ttaaaattgg atttgtaata ataaaacgca
     1261 attgtttgtt attgtggcgc tctatcatag atgtcgctat aaacctattc agcacaatat
     1321 attgttttca ttttaatatt gtacatataa gtagtagggt acaatcagta aattgaacgg
     1381 agaatattat tcataaaaat acgatagtaa cgggtgatat attcattaga atgaaccgaa
     1441 accggcggta aggatctgag ctacacatgc tcaggttttt tacaacgtgc acaacagaat
     1501 tgaaagcaaa tatcatgcga tcataggctt ctcgcatatc tcattaaagc aggacaagct
     1561 caagaagtca ccattgcaga gattccgtac accaccaatc cagtgatagc agctccgaac
     1621 acgaaagtca tgatctttcc gaactttgaa gatttctcag ggttcttagc gatctcgtta
     1681 gccacaccct cagtaagctt agccacctgt gcctcgtcca cgttcttaag ctgtccagat
     1741 gccttcaact tctggaaagc tgattgccac ttgttatttg ttgtgtccaa gtttgtaagc
     1801 tttcctgcct ggatcttagc cactgcagcc tgccacttgt tctggtcagc cacagggatc
     1861 ttgatgtcgt cgatgttctt gaagttgtta gccttcaact tttcaagagc agattgccat
     1921 ttgttttttc cagcagtagt agtctcagcc accttctcag cgttctgcaa agccttctgt
     1981 gccttaagct tttccaatgc ctgttgccac ttatttccac cagcagcagc cacagcatca
     2041 tcaagctgtc ctgctccctt gatcttagcc aatgcgtcgt ccacctgtcc cactcccttc
     2101 atcttctcca aagcgtcatc caattgacct gccttgaact ttgatgccaa gtctgcccat
     2161 cctgccattg tatcgataat tgtaaatgta attgtaatgt tgtttgttgt ttgttgttgt
     2221 tggtaattgt tgtaaaaatg agctcttata ctcgagcgtg tcctctccaa atgaaatgaa
     2281 cttccttata tagaggaagg gtcttgcgaa ggatagtggg attgtgcgtc atcccttacg
     2341 tcagtggaga tgtcacatca atccacttgc tttgtagacg tggttggaac ctcttctttt
     2401 tccacgatgc tcctcgtggg tgggggtcca tctttgggac cactgtcggc agagagatct
     2461 tgaatgatag cctttccttt atcgcaatga tggcatttgt aggagccacc ttccttttct
     2521 actgtccttt cgatgaagtg acagatagct gggcaatgga atccgaggag gtttcccgaa
     2581 attatccttt gttgaaaagt ctcaatagcc ctttgatctt ctgagactgt atctttgaca
     2641 tttttggagt agaccagagt gtcgtgctcc accatgttga cgaagatttt cttcttgtca
     2701 ttgagtcgta aaagactctg tatgaactgt tcgccagtct tgacggcgag ttctgttaga
     2761 tcctcgattt gaatcttaga ctccatgcat ggccttagat tcagtaggaa ctaccttttt
     2821 agagactcca atctctatta cttgccttgg tttatgaagc aagccttgaa tcgtccatac
     2881 tggaatagta cttctgatct tgagaaatat gtctttctct gtgttcttga tgcaattagt
     2941 cctgaatctt ttgactgcat ctttaacctt cttgggaagg tatttgatct cctggagatt
     3001 gttactcggg tagatcgtct tgatgagact tgctgcgtag gcctctctaa ccatctgtgg
     3061 gtcagcattc tttctgaaat tgaagaggct aaccttctca ttatcagtgg tgaacatagt
     3121 gtcgtcacct tcaccttcga acttccttcc tagatcgtaa agatagagga aatcgtccat
     3181 tgtaatctcc ggggcaaagg agatctcttt tggggctgga tcactgctgg gccttttggt
     3241 tcctagcgtg agccagtggg ctttttgctt tggtgggctt gttagggcct tagcaaagct
     3301 cttgggcttg agttgagctt ctcctttggg gatgaagttc aacctgtctg tttgctgact
     3361 tgttgtgtac gcgtcagctg ctgctcttgc ctctgtaata gtggcaaatt tcttgtgtgc
     3421 aactccggga acgccgtttg ttgccgcctt tgtacaaccc cagtcatcgt atataccggc
     3481 atgtggaccg ttatacacaa cgtagtagtt gatatgaggg tgttgaatac ccgattctgc
     3541 tctgagagga gcaactgtgc tgttaagctc agatttttgt gggattggaa ttcctccaag
     3601 cttgaattct tgcagcgctc caattcagca agcgtcgatc tagtaacata gatgacaccg
     3661 cgcgcgataa tttatcctag tttgcgcgct atattttgtt ttctatcgcg tattaaatgt
     3721 ataattgcgg gactctaatc ataaaaaccc atctcataaa taacgtcatg cattacatgt
     3781 taattattac atgcttaacg taattcaaca gaaattatat gataatcatc gcaagaccgg
     3841 caacaggatt caatcttaag aaactttatt gccaaatgtt tgaacgatct gcttgactct
     3901 agatccagag tcccgctcaa atctcagtca caggaagaac aggtctagga ggaacaggaa
     3961 gagagaaatc aagctgccag aatcccacat catgccaatt tccatgtttg aaaccagcag
     4021 ctctaagcat tcccctagga gcataaccaa gagcctcatg catcctcaca gaaggatcat
     4081 taggaagtcc aatcacagca accacagact tgaaaccctg agcctccaaa gacttcaaaa
     4141 gatgagtgta aagagtagat ccaagaccag ttctttgatg cctaggagac acgtaaacag
     4201 tagactcagc agtccaatcg taagcattcc tagccttcca aggaccagca taagcaatac
     4261 cagccacttc accatccact tcagcaacaa gccaaggata cctctctcta agcctcacaa
     4321 gatcatcagt ccactcttgt ggttcttgag gctcagtcct gaaattcaca gtagaagtct
     4381 caatgtaatg attcacaata gtgcacacag caggcatatc agcctcagta gcccttctaa
     4441 tatcagcagg ccttctttca ggagacatgc gaaacgatcc agatccggtg cagattattt
     4501 ggattgagag tgaatatgag actctaattg gataccgagg ggaatttatg gaacgtcagt
     4561 ggagcatttt gacaagaaat atttgctagc tgatagtgac cttaggcgac ttttgaacgc
     4621 gcaataatgg tttctgacgt atgtgcttag ctcattaaac tccagaaacc cgcggctgag
     4681 tggctccttc aacgttgcgg ttctgtcagt tccaaacgta aaacggcttg tcccgcgtca
     4741 tcggcggggg tcataacgtg actcccttaa ttctccgctc cggatccgaa ttcggcacag
     4801 agtgttcaac cccagtacat taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc
     4861 aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc cgaccggcag
     4921 ctcggcacaa aatcaccact cgatacaggc agcccatcaa ccatgactac tgtccggtcc
     4981 aacagatccg agcaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     5041 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     5101 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     5161 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     5221 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     5281 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5341 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5401 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5461 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5521 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5581 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     5641 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     5701 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     5761 atatgagtaa acttggtctg acagctaaaa caattcatcc agtaaaatat aatattttat
     5821 tttctcccaa tcaggcttga tccccagtaa gtcaaaaaat agctcgacat actgttcttc
     5881 cccgatatcc tccctgagcg accggacgca gaaggcaatg tcataccact tgtccgccct
     5941 gccgcttctc ccaagatcaa taaagccact tactttgcca tctttcacaa agatgttgct
     6001 gtctcccagg tcgccgtggg aaaagacaag ttcctcttcg ggcttttccg tctttaaaaa
     6061 atcatacagc tcgcgcggat ctttaaatgg agtgtcctct tcccagtttt cgcaatccac
     6121 atcggccaga tcgttattca gtaagtaatc caattcggct aagcggctgt ctaagctatt
     6181 cgtataggga caatccgata tgtcgatgga gtgaaagagc ctgatgcact ccgcatacag
     6241 ctcgataatc ttttcagggc tttgttcatc ttcatactct tccgagcaaa ggacgccatc
     6301 ggcctcactc atgagcagat tgctccagcc atcatgccgt tcaaagtgca ggacctttgg
     6361 aacaggcagc tttccttcca gccatagcat catgtccttt tcccgttcca catcataggt
     6421 ggtcccttta taccggctgt ccgtcatttt taaatatagg ttttcatttt ctcccaccag
     6481 cttatatacc ttagcaggag acattccttc cgtatctttt acgcagcggt atttttcgat
     6541 cagttttttc aattccggtg atattctcat tttagccata ctcttccttt ttcaatatta
     6601 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6661 aaataaacaa ataggggttc cgcgcactcg gagccttgat gtgggcgctt tttgcagctg
     6721 ttcggctgtg cgctggccag acagttatgc acaggccagg cgggttttaa gagttttaat
     6781 aagttttaaa gagttttagg cggaaaaatc gccttttttc tcttttatat cagtcactta
     6841 catgtgtgac cggttcccaa tgtacggctt tgggttccca atgtacgggt tccggttccc
     6901 aatgtacggc tttgggttcc caatgtacgt gctatccaca ggaaagaagc tttttcgacc
     6961 tttttcccct gctagggcaa tttgccctag catctgctcc gtacattagg aaccggcgga
     7021 tgcttcgccc tcgatcaggt tgcggtagcg catgactagg atcgggccag cctgccccgc
     7081 ctcctccttc aaatcgtact ccggcaggtc atttgacccg atcagcttgc gcacggtgaa
     7141 acagaacttc ttgaactctc cggcgctgcc actgcgttcg tagatcgtct tgaacaacca
     7201 tctggcttct gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc
     7261 gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg tccgggttct tgccttctgt
     7321 gatctcgcgg tacatccaat cagctagctc gatctcgatg tactccggcc gcccggtttc
     7381 gctctttacg atcttgtagc ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg
     7441 gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta accgaatgca
     7501 ggtttctacc aggtcgtctt tctgctttcc gccatcggct cgccggcaga acttgagtac
     7561 gtccgcaacg tgtggacgga acacgcggcc gggcttgtct cccttccctt cccggtatcg
     7621 gttcatggat tcggttagat gggaaaccgc catcagtacc aggtcgtaat cccacacact
     7681 ggccatgccg gccggccctg cggaaacctc tacgtgcccg tctggaagct cgtagcggat
     7741 cacctcgcca gctcgtcggt cacgcttcga cagacggaaa acggccacgt ccatgatgct
     7801 gcgactatcg cgggtgccca cgtcatagag catcggaacg aaaaaatctg gttgctcgtc
     7861 gcccttgggc ggcttcctaa tcgacgacgc accggctgcc ggcggttgcc gggattcttt
     7921 gcggattcga tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg
     7981 ttgccgctgg gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc
     8041 gccgatttgt accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg
     8101 gttccagtgc cattgcaggg ccggcagaca acccagccgc ttacgcctgg ccaaccgccc
     8161 gttcctccac acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg
     8221 ccgcctcctt tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg
     8281 tagctgcgcg atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta
     8341 catcttcagc ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg
     8401 cgtgtctgcc aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc
     8461 acttagcgtg tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt
     8521 gatttaattt cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat
     8581 tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg
     8641 gactcaagaa tgggcagctc gtacccggcc agcgcctcgg caacctcacc gc
//
