LOCUS       sequence-342137-        8233 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 8233)
  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 8233)
  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_04089
FEATURES             Location/Qualifiers
     source          1..8233
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    626..650
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(717..1424)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1672..2017)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1709..1729)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3050..3302)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(3902..4084)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(3938..4116)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4279..4303
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4439..5027)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4519..4538)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5198..5992)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(5993..6097)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6159..6353
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6419..7492)
                     /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(7921..8233,1..317))
                     /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 tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc
       61 tggggcgctc cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca
      121 atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa
      181 tcgcgggcac tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc
      241 agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt
      301 acagcgataa ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat
      361 acgcagcgac cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc
      421 ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca
      481 aaccggccag gatttcatgc agccgcacgg ctgactcccg aggggaaccc tgtggttggc
      541 atgcacatac aaatggacga acggataaac cttttcacgc ccttttaaat atccgattat
      601 tctaataaac gctcttttct cttaggttta cccgccaata tatcctgtca aacactgata
      661 gtttaaacgg atcacttcgt gtccctcggt cacatgtgca tcctctagta gcggtcctgc
      721 tgagcctcga catgttgtcg caaaattcgc cctggacccg cccaacgatt tgtcgtcact
      781 gtcaaggttt gacctgcact tcatttgggg cccacataca ccaaaaaaat gctgcataat
      841 tctcggggca gcaagtcggt tacccggccg ccgtgctgga ccgggttgaa tggtgcccgt
      901 aactttcggt agagcggacg gccaatactc aacttcaagg aatctcaccc atgcgcgccg
      961 gcggggaacc ggagttccct tcagtgagcg ttattagttc gccgctcggt gtgtcgtaga
     1021 tactagcccc tggggcactt ttgaaatttg aataagattt atgtaatcag tcttttaggt
     1081 ttgaccggtt ctgccgcttt ttttaaaatt ggatttgtaa taataaaacg caattgtttg
     1141 ttattgtggc gctctatcat agatgtcgct ataaacctat tcagcacaat atattgtttt
     1201 cattttaata ttgtacatat aagtagtagg gtacaatcag taaattgaac ggagaatatt
     1261 attcataaaa atacgatagt aacgggtgat atattcatta gaatgaaccg aaaccggcgg
     1321 taaggatctg agctacacat gctcaggttt tttacaacgt gcacaacaga attgaaagca
     1381 aatatcatgc gatcataggc ttctcgcata tctcattaaa gcaggacaag ctcacttagc
     1441 aggcttcttg atcaccttag acaacttagc catcttcttc tcaagaagag ccttcttagc
     1501 agcagcgatc aacttagaca tgtccttctc agcaggcctc ttgaagatct tgttagtaga
     1561 ctgagcagtc accctcacca ttgtatcgat aattgtaaat gtaattgtaa tgttgtttgt
     1621 tgtttgttgt tgttggtaat tgttgtaaaa atgagctctt atactcgagc gtgtcctctc
     1681 caaatgaaat gaacttcctt atatagagga agggtcttgc gaaggatagt gggattgtgc
     1741 gtcatccctt acgtcagtgg agatgtcaca tcaatccact tgctttgtag acgtggttgg
     1801 aacctcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc
     1861 ggcagagaga tcttgaatga tagcctttcc tttatcgcaa tgatggcatt tgtaggagcc
     1921 accttccttt tctactgtcc tttcgatgaa gtgacagata gctgggcaat ggaatccgag
     1981 gaggtttccc gaaattatcc tttgttgaaa agtctcaata gccctttgat cttctgagac
     2041 tgtatctttg acatttttgg agtagaccag agtgtcgtgc tccaccatgt tgacgaagat
     2101 tttcttcttg tcattgagtc gtaaaagact ctgtatgaac tgttcgccag tcttgacggc
     2161 gagttctgtt agatcctcga tttgaatctt agactccatg catggcctta gattcagtag
     2221 gaactacctt tttagagact ccaatctcta ttacttgcct tggtttatga agcaagcctt
     2281 gaatcgtcca tactggaata gtacttctga tcttgagaaa tatgtctttc tctgtgttct
     2341 tgatgcaatt agtcctgaat cttttgactg catctttaac cttcttggga aggtatttga
     2401 tctcctggag attgttactc gggtagatcg tcttgatgag acttgctgcg taggcctctc
     2461 taaccatctg tgggtcagca ttctttctga aattgaagag gctaaccttc tcattatcag
     2521 tggtgaacat agtgtcgtca ccttcacctt cgaacttcct tcctagatcg taaagataga
     2581 ggaaatcgtc cattgtaatc tccggggcaa aggagatctc ttttggggct ggatcactgc
     2641 tgggcctttt ggttcctagc gtgagccagt gggctttttg ctttggtggg cttgttaggg
     2701 ccttagcaaa gctcttgggc ttgagttgag cttctccttt ggggatgaag ttcaacctgt
     2761 ctgtttgctg acttgttgtg tacgcgtcag ctgctgctct tgcctctgta atagtggcaa
     2821 atttcttgtg tgcaactccg ggaacgccgt ttgttgccgc ctttgtacaa ccccagtcat
     2881 cgtatatacc ggcatgtgga ccgttataca caacgtagta gttgatatga gggtgttgaa
     2941 tacccgattc tgctctgaga ggagcaactg tgctgttaag ctcagatttt tgtgggattg
     3001 gaattcctcc aagcttgaat tcttgcagcg ctccaattca gcaagcgtcg atctagtaac
     3061 atagatgaca ccgcgcgcga taatttatcc tagtttgcgc gctatatttt gttttctatc
     3121 gcgtattaaa tgtataattg cgggactcta atcataaaaa cccatctcat aaataacgtc
     3181 atgcattaca tgttaattat tacatgctta acgtaattca acagaaatta tatgataatc
     3241 atcgcaagac cggcaacagg attcaatctt aagaaacttt attgccaaat gtttgaacga
     3301 tctgcttgac tctagatcca gagtcccgct caaatctcag tcacaggaag aacaggtcta
     3361 ggaggaacag gaagagagaa atcaagctgc cagaatccca catcatgcca atttccatgt
     3421 ttgaaaccag cagctctaag cattccccta ggagcataac caagagcctc atgcatcctc
     3481 acagaaggat cattaggaag tccaatcaca gcaaccacag acttgaaacc ctgagcctcc
     3541 aaagacttca aaagatgagt gtaaagagta gatccaagac cagttctttg atgcctagga
     3601 gacacgtaaa cagtagactc agcagtccaa tcgtaagcat tcctagcctt ccaaggacca
     3661 gcataagcaa taccagccac ttcaccatcc acttcagcaa caagccaagg atacctctct
     3721 ctaagcctca caagatcatc agtccactct tgtggttctt gaggctcagt cctgaaattc
     3781 acagtagaag tctcaatgta atgattcaca atagtgcaca cagcaggcat atcagcctca
     3841 gtagcccttc taatatcagc aggccttctt tcaggagaca tgcgaaacga tccagatccg
     3901 gtgcagatta tttggattga gagtgaatat gagactctaa ttggataccg aggggaattt
     3961 atggaacgtc agtggagcat tttgacaaga aatatttgct agctgatagt gaccttaggc
     4021 gacttttgaa cgcgcaataa tggtttctga cgtatgtgct tagctcatta aactccagaa
     4081 acccgcggct gagtggctcc ttcaacgttg cggttctgtc agttccaaac gtaaaacggc
     4141 ttgtcccgcg tcatcggcgg gggtcataac gtgactccct taattctccg ctccggatcc
     4201 gaattcggca cagagtgttc aaccccagta cattaaaaac gtccgcaatg tgttattaag
     4261 ttgtctaagc gtcaatttgt ttacaccaca atatatcctg ccaccagcca gccaacagct
     4321 ccccgaccgg cagctcggca caaaatcacc actcgataca ggcagcccat caaccatgac
     4381 tactgtccgg tccaacagat ccgagcagga accgtaaaaa ggccgcgttg ctggcgtttt
     4441 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4501 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4561 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4621 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4681 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4741 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4801 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4861 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4921 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4981 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5041 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5101 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5161 caatctaaag tatatatgag taaacttggt ctgacagcta aaacaattca tccagtaaaa
     5221 tataatattt tattttctcc caatcaggct tgatccccag taagtcaaaa aatagctcga
     5281 catactgttc ttccccgata tcctccctga gcgaccggac gcagaaggca atgtcatacc
     5341 acttgtccgc cctgccgctt ctcccaagat caataaagcc acttactttg ccatctttca
     5401 caaagatgtt gctgtctccc aggtcgccgt gggaaaagac aagttcctct tcgggctttt
     5461 ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa tggagtgtcc tcttcccagt
     5521 tttcgcaatc cacatcggcc agatcgttat tcagtaagta atccaattcg gctaagcggc
     5581 tgtctaagct attcgtatag ggacaatccg atatgtcgat ggagtgaaag agcctgatgc
     5641 actccgcata cagctcgata atcttttcag ggctttgttc atcttcatac tcttccgagc
     5701 aaaggacgcc atcggcctca ctcatgagca gattgctcca gccatcatgc cgttcaaagt
     5761 gcaggacctt tggaacaggc agctttcctt ccagccatag catcatgtcc ttttcccgtt
     5821 ccacatcata ggtggtccct ttataccggc tgtccgtcat ttttaaatat aggttttcat
     5881 tttctcccac cagcttatat accttagcag gagacattcc ttccgtatct tttacgcagc
     5941 ggtatttttc gatcagtttt ttcaattccg gtgatattct cattttagcc atactcttcc
     6001 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     6061 aatgtattta gaaaaataaa caaatagggg ttccgcgcac tcggagcctt gatgtgggcg
     6121 ctttttgcag ctgttcggct gtgcgctggc cagacagtta tgcacaggcc aggcgggttt
     6181 taagagtttt aataagtttt aaagagtttt aggcggaaaa atcgcctttt ttctctttta
     6241 tatcagtcac ttacatgtgt gaccggttcc caatgtacgg ctttgggttc ccaatgtacg
     6301 ggttccggtt cccaatgtac ggctttgggt tcccaatgta cgtgctatcc acaggaaaga
     6361 agctttttcg acctttttcc cctgctaggg caatttgccc tagcatctgc tccgtacatt
     6421 aggaaccggc ggatgcttcg ccctcgatca ggttgcggta gcgcatgact aggatcgggc
     6481 cagcctgccc cgcctcctcc ttcaaatcgt actccggcag gtcatttgac ccgatcagct
     6541 tgcgcacggt gaaacagaac ttcttgaact ctccggcgct gccactgcgt tcgtagatcg
     6601 tcttgaacaa ccatctggct tctgccttgc ctgcggcgcg gcgtgccagg cggtagagaa
     6661 aacggccgat gccgggatcg atcaaaaagt aatcggggtg aaccgtcagc acgtccgggt
     6721 tcttgccttc tgtgatctcg cggtacatcc aatcagctag ctcgatctcg atgtactccg
     6781 gccgcccggt ttcgctcttt acgatcttgt agcggctaat caaggcttca ccctcggata
     6841 ccgtcaccag gcggccgttc ttggccttct tcgtacgctg catggcaacg tgcgtggtgt
     6901 ttaaccgaat gcaggtttct accaggtcgt ctttctgctt tccgccatcg gctcgccggc
     6961 agaacttgag tacgtccgca acgtgtggac ggaacacgcg gccgggcttg tctcccttcc
     7021 cttcccggta tcggttcatg gattcggtta gatgggaaac cgccatcagt accaggtcgt
     7081 aatcccacac actggccatg ccggccggcc ctgcggaaac ctctacgtgc ccgtctggaa
     7141 gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt cgacagacgg aaaacggcca
     7201 cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata gagcatcgga acgaaaaaat
     7261 ctggttgctc gtcgcccttg ggcggcttcc taatcgacga cgcaccggct gccggcggtt
     7321 gccgggattc tttgcggatt cgatcagcgg ccgcttgcca cgattcaccg gggcgtgctt
     7381 ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt caacttctcc accaggtcat
     7441 cacccagcgc cgcgccgatt tgtaccgggc cggatggttt gcgaccgctc acgccgattc
     7501 ctcgggcttg ggggttccag tgccattgca gggccggcag acaacccagc cgcttacgcc
     7561 tggccaaccg cccgttcctc cacacatggg gcattccacg gcgtcggtgc ctggttgttc
     7621 ttgattttcc atgccgcctc ctttagccgc taaaattcat ctactcattt attcatttgc
     7681 tcatttactc tggtagctgc gcgatgtatt cagatagcag ctcggtaatg gtcttgcctt
     7741 ggcgtaccgc gtacatcttc agcttggtgt gatcctccgc cggcaactga aagttgaccc
     7801 gcttcatggc tggcgtgtct gccaggctgg ccaacgttgc agccttgctg ctgcgtgcgc
     7861 tcggacggcc ggcacttagc gtgtttgtgc ttttgctcat tttctcttta cctcattaac
     7921 tcaaatgagt tttgatttaa tttcagcggc cagcgcctgg acctcgcggg cagcgtcgcc
     7981 ctcgggttct gattcaagaa cggttgtgcc ggcggcggca gtgcctgggt agctcacgcg
     8041 ctgcgtgata cgggactcaa gaatgggcag ctcgtacccg gccagcgcct cggcaacctc
     8101 accgccgatg cgcgtgcctt tgatcgcccg cgacacgaca aaggccgctt gtagccttcc
     8161 atccgtgacc tcaatgcgct gcttaaccag ctccaccagg tcggcggtgg cccatatgtc
     8221 gtaagggctt ggc
//
