LOCUS       sequence-342201-        8386 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 8386)
  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 8386)
  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_09160
FEATURES             Location/Qualifiers
     source          1..8386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    621..645
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(712..1419)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1820..2165)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1857..1877)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3198..3450)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4050..4232)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4086..4264)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4427..4451
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4587..5175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4667..4686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5346..6140)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6141..6245)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6307..6501
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6567..7640)
                     /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(8069..8386,1..312))
                     /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 cggaatcagc acgaagtcgg ctgccttgat cgcggacaca gccaagtccg ccgcctgggg
       61 cgctccgtcg atcactacga agtcgcgccg gccgatggcc ttcacgtcgc ggtcaatcgt
      121 cgggcggtcg atgccgacaa cggttagcgg ttgatcttcc cgcacggccg cccaatcgcg
      181 ggcactgccc tggggatcgg aatcgactaa cagaacatcg gccccggcga gttgcagggc
      241 gcgggctaga tgggttgcga tggtcgtctt gcctgacccg cctttctggt taagtacagc
      301 gataaccttc atgcgttccc cttgcgtatt tgtttattta ctcatcgcat catatacgca
      361 gcgaccgcat gacgcaagct gttttactca aatacacatc acctttttag acggcggcgc
      421 tcggtttctt cagcggccaa gctggccggc caggccgcca gcttggcatc agacaaaccg
      481 gccaggattt catgcagccg cacggctgac tcccgagggg aaccctgtgg ttggcatgca
      541 catacaaatg gacgaacgga taaacctttt cacgcccttt taaatatccg attattctaa
      601 taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac tgatagttta
      661 aacggatcac ttcgtgtccc tcggtcacat gtgcatcctc tagtagcggt cctgctgagc
      721 ctcgacatgt tgtcgcaaaa ttcgccctgg acccgcccaa cgatttgtcg tcactgtcaa
      781 ggtttgacct gcacttcatt tggggcccac atacaccaaa aaaatgctgc ataattctcg
      841 gggcagcaag tcggttaccc ggccgccgtg ctggaccggg ttgaatggtg cccgtaactt
      901 tcggtagagc ggacggccaa tactcaactt caaggaatct cacccatgcg cgccggcggg
      961 gaaccggagt tcccttcagt gagcgttatt agttcgccgc tcggtgtgtc gtagatacta
     1021 gcccctgggg cacttttgaa atttgaataa gatttatgta atcagtcttt taggtttgac
     1081 cggttctgcc gcttttttta aaattggatt tgtaataata aaacgcaatt gtttgttatt
     1141 gtggcgctct atcatagatg tcgctataaa cctattcagc acaatatatt gttttcattt
     1201 taatattgta catataagta gtagggtaca atcagtaaat tgaacggaga atattattca
     1261 taaaaatacg atagtaacgg gtgatatatt cattagaatg aaccgaaacc ggcggtaagg
     1321 atctgagcta cacatgctca ggttttttac aacgtgcaca acagaattga aagcaaatat
     1381 catgcgatca taggcttctc gcatatctca ttaaagcagg acaagctcaa gccttgttgt
     1441 tctgctcctt cttgatcaag aacttcacgt acctcttcac agtcttaagc ctctcagcgt
     1501 cagtgaactg cttgtctccc ttcaacatct tcaacataga agcccttccc attcccacag
     1561 cgtcagcgaa agcgaagatc ttgtccacct tgtcgtccac cattgcagcc acgtgcctga
     1621 tcacagccaa gttagaagtc ttttccaact tgtccaaagg agaaagagac ttagtcttga
     1681 acacctggtc caacacctta gcagcagcct ccttctgagc gttcaccatt gtatcgataa
     1741 ttgtaaatgt aattgtaatg ttgtttgttg tttgttgttg ttggtaattg ttgtaaaaat
     1801 gagctcttat actcgagcgt gtcctctcca aatgaaatga acttccttat atagaggaag
     1861 ggtcttgcga aggatagtgg gattgtgcgt catcccttac gtcagtggag atgtcacatc
     1921 aatccacttg ctttgtagac gtggttggaa cctcttcttt ttccacgatg ctcctcgtgg
     1981 gtgggggtcc atctttggga ccactgtcgg cagagagatc ttgaatgata gcctttcctt
     2041 tatcgcaatg atggcatttg taggagccac cttccttttc tactgtcctt tcgatgaagt
     2101 gacagatagc tgggcaatgg aatccgagga ggtttcccga aattatcctt tgttgaaaag
     2161 tctcaatagc cctttgatct tctgagactg tatctttgac atttttggag tagaccagag
     2221 tgtcgtgctc caccatgttg acgaagattt tcttcttgtc attgagtcgt aaaagactct
     2281 gtatgaactg ttcgccagtc ttgacggcga gttctgttag atcctcgatt tgaatcttag
     2341 actccatgca tggccttaga ttcagtagga actacctttt tagagactcc aatctctatt
     2401 acttgccttg gtttatgaag caagccttga atcgtccata ctggaatagt acttctgatc
     2461 ttgagaaata tgtctttctc tgtgttcttg atgcaattag tcctgaatct tttgactgca
     2521 tctttaacct tcttgggaag gtatttgatc tcctggagat tgttactcgg gtagatcgtc
     2581 ttgatgagac ttgctgcgta ggcctctcta accatctgtg ggtcagcatt ctttctgaaa
     2641 ttgaagaggc taaccttctc attatcagtg gtgaacatag tgtcgtcacc ttcaccttcg
     2701 aacttccttc ctagatcgta aagatagagg aaatcgtcca ttgtaatctc cggggcaaag
     2761 gagatctctt ttggggctgg atcactgctg ggccttttgg ttcctagcgt gagccagtgg
     2821 gctttttgct ttggtgggct tgttagggcc ttagcaaagc tcttgggctt gagttgagct
     2881 tctcctttgg ggatgaagtt caacctgtct gtttgctgac ttgttgtgta cgcgtcagct
     2941 gctgctcttg cctctgtaat agtggcaaat ttcttgtgtg caactccggg aacgccgttt
     3001 gttgccgcct ttgtacaacc ccagtcatcg tatataccgg catgtggacc gttatacaca
     3061 acgtagtagt tgatatgagg gtgttgaata cccgattctg ctctgagagg agcaactgtg
     3121 ctgttaagct cagatttttg tgggattgga attcctccaa gcttgaattc ttgcagcgct
     3181 ccaattcagc aagcgtcgat ctagtaacat agatgacacc gcgcgcgata atttatccta
     3241 gtttgcgcgc tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat
     3301 cataaaaacc catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac
     3361 gtaattcaac agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa
     3421 gaaactttat tgccaaatgt ttgaacgatc tgcttgactc tagatccaga gtcccgctca
     3481 aatctcagtc acaggaagaa caggtctagg aggaacagga agagagaaat caagctgcca
     3541 gaatcccaca tcatgccaat ttccatgttt gaaaccagca gctctaagca ttcccctagg
     3601 agcataacca agagcctcat gcatcctcac agaaggatca ttaggaagtc caatcacagc
     3661 aaccacagac ttgaaaccct gagcctccaa agacttcaaa agatgagtgt aaagagtaga
     3721 tccaagacca gttctttgat gcctaggaga cacgtaaaca gtagactcag cagtccaatc
     3781 gtaagcattc ctagccttcc aaggaccagc ataagcaata ccagccactt caccatccac
     3841 ttcagcaaca agccaaggat acctctctct aagcctcaca agatcatcag tccactcttg
     3901 tggttcttga ggctcagtcc tgaaattcac agtagaagtc tcaatgtaat gattcacaat
     3961 agtgcacaca gcaggcatat cagcctcagt agcccttcta atatcagcag gccttctttc
     4021 aggagacatg cgaaacgatc cagatccggt gcagattatt tggattgaga gtgaatatga
     4081 gactctaatt ggataccgag gggaatttat ggaacgtcag tggagcattt tgacaagaaa
     4141 tatttgctag ctgatagtga ccttaggcga cttttgaacg cgcaataatg gtttctgacg
     4201 tatgtgctta gctcattaaa ctccagaaac ccgcggctga gtggctcctt caacgttgcg
     4261 gttctgtcag ttccaaacgt aaaacggctt gtcccgcgtc atcggcgggg gtcataacgt
     4321 gactccctta attctccgct ccggatccga attcggcaca gagtgttcaa ccccagtaca
     4381 ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt acaccacaat
     4441 atatcctgcc accagccagc caacagctcc ccgaccggca gctcggcaca aaatcaccac
     4501 tcgatacagg cagcccatca accatgacta ctgtccggtc caacagatcc gagcaggaac
     4561 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4621 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4681 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4741 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4801 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4861 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4921 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4981 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5041 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5101 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5161 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5221 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5281 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5341 gacagctaaa acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg
     5401 atccccagta agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgagc
     5461 gaccggacgc agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca
     5521 ataaagccac ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg
     5581 gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga
     5641 tctttaaatg gagtgtcctc ttcccagttt tcgcaatcca catcggccag atcgttattc
     5701 agtaagtaat ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat
     5761 atgtcgatgg agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg
     5821 ctttgttcat cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga
     5881 ttgctccagc catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc
     5941 agccatagca tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg
     6001 tccgtcattt ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga
     6061 gacattcctt ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt
     6121 gatattctca ttttagccat actcttcctt tttcaatatt attgaagcat ttatcagggt
     6181 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     6241 ccgcgcactc ggagccttga tgtgggcgct ttttgcagct gttcggctgt gcgctggcca
     6301 gacagttatg cacaggccag gcgggtttta agagttttaa taagttttaa agagttttag
     6361 gcggaaaaat cgcctttttt ctcttttata tcagtcactt acatgtgtga ccggttccca
     6421 atgtacggct ttgggttccc aatgtacggg ttccggttcc caatgtacgg ctttgggttc
     6481 ccaatgtacg tgctatccac aggaaagaag ctttttcgac ctttttcccc tgctagggca
     6541 atttgcccta gcatctgctc cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg
     6601 ttgcggtagc gcatgactag gatcgggcca gcctgccccg cctcctcctt caaatcgtac
     6661 tccggcaggt catttgaccc gatcagcttg cgcacggtga aacagaactt cttgaactct
     6721 ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc atctggcttc tgccttgcct
     6781 gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa
     6841 tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg gtacatccaa
     6901 tcagctagct cgatctcgat gtactccggc cgcccggttt cgctctttac gatcttgtag
     6961 cggctaatca aggcttcacc ctcggatacc gtcaccaggc ggccgttctt ggccttcttc
     7021 gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac caggtcgtct
     7081 ttctgctttc cgccatcggc tcgccggcag aacttgagta cgtccgcaac gtgtggacgg
     7141 aacacgcggc cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga
     7201 tgggaaaccg ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct
     7261 gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg
     7321 tcacgcttcg acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc
     7381 acgtcataga gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta
     7441 atcgacgacg caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc
     7501 gcttgccacg attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc
     7561 gcggccttca acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg
     7621 gatggtttgc gaccgctcac gccgattcct cgggcttggg ggttccagtg ccattgcagg
     7681 gccggcagac aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatggggc
     7741 attccacggc gtcggtgcct ggttgttctt gattttccat gccgcctcct ttagccgcta
     7801 aaattcatct actcatttat tcatttgctc atttactctg gtagctgcgc gatgtattca
     7861 gatagcagct cggtaatggt cttgccttgg cgtaccgcgt acatcttcag cttggtgtga
     7921 tcctccgccg gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc caggctggcc
     7981 aacgttgcag ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt gtttgtgctt
     8041 ttgctcattt tctctttacc tcattaactc aaatgagttt tgatttaatt tcagcggcca
     8101 gcgcctggac ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg gttgtgccgg
     8161 cggcggcagt gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga atgggcagct
     8221 cgtacccggc cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg atcgcccgcg
     8281 acacgacaaa ggccgcttgt agccttccat ccgtgacctc aatgcgctgc ttaaccagct
     8341 ccaccaggtc ggcggtggcc catatgtcgt aagggcttgg ctgcac
//
