LOCUS       sequence-342798-        9280 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 9280)
  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 9280)
  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_22922
FEATURES             Location/Qualifiers
     source          1..9280
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    647..671
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(738..1445)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2740..3085)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2777..2797)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4118..4370)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4970..5152)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5006..5184)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5347..5371
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5507..6095)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5587..5606)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6266..7060)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7061..7165)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7227..7421
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7487..8560)
                     /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(8989..9280,1..338))
                     /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 catatgtcgt aagggcttgg ctgcaccgga atcagcacga agtcggctgc cttgatcgcg
       61 gacacagcca agtccgccgc ctggggcgct ccgtcgatca ctacgaagtc gcgccggccg
      121 atggccttca cgtcgcggtc aatcgtcggg cggtcgatgc cgacaacggt tagcggttga
      181 tcttcccgca cggccgccca atcgcgggca ctgccctggg gatcggaatc gactaacaga
      241 acatcggccc cggcgagttg cagggcgcgg gctagatggg ttgcgatggt cgtcttgcct
      301 gacccgcctt tctggttaag tacagcgata accttcatgc gttccccttg cgtatttgtt
      361 tatttactca tcgcatcata tacgcagcga ccgcatgacg caagctgttt tactcaaata
      421 cacatcacct ttttagacgg cggcgctcgg tttcttcagc ggccaagctg gccggccagg
      481 ccgccagctt ggcatcagac aaaccggcca ggatttcatg cagccgcacg gctgactccc
      541 gaggggaacc ctgtggttgg catgcacata caaatggacg aacggataaa ccttttcacg
      601 cccttttaaa tatccgatta ttctaataaa cgctcttttc tcttaggttt acccgccaat
      661 atatcctgtc aaacactgat agtttaaacg gatcacttcg tgtccctcgg tcacatgtgc
      721 atcctctagt agcggtcctg ctgagcctcg acatgttgtc gcaaaattcg ccctggaccc
      781 gcccaacgat ttgtcgtcac tgtcaaggtt tgacctgcac ttcatttggg gcccacatac
      841 accaaaaaaa tgctgcataa ttctcggggc agcaagtcgg ttacccggcc gccgtgctgg
      901 accgggttga atggtgcccg taactttcgg tagagcggac ggccaatact caacttcaag
      961 gaatctcacc catgcgcgcc ggcggggaac cggagttccc ttcagtgagc gttattagtt
     1021 cgccgctcgg tgtgtcgtag atactagccc ctggggcact tttgaaattt gaataagatt
     1081 tatgtaatca gtcttttagg tttgaccggt tctgccgctt tttttaaaat tggatttgta
     1141 ataataaaac gcaattgttt gttattgtgg cgctctatca tagatgtcgc tataaaccta
     1201 ttcagcacaa tatattgttt tcattttaat attgtacata taagtagtag ggtacaatca
     1261 gtaaattgaa cggagaatat tattcataaa aatacgatag taacgggtga tatattcatt
     1321 agaatgaacc gaaaccggcg gtaaggatct gagctacaca tgctcaggtt ttttacaacg
     1381 tgcacaacag aattgaaagc aaatatcatg cgatcatagg cttctcgcat atctcattaa
     1441 agcaggacaa gctcagtgtg tgtgctgcaa gtacctctgg taatatttat agttagttgc
     1501 ctccttaaga gcagtctcgt ccaaaccgtc aagtcccaaa gagaacttca cgtaccactc
     1561 aggcctctcc ttcttagccc agataagagc cttctcgtta agctccctac cagaagcctc
     1621 cttgttcaca aggttaggca attttgtttc tcccttgttg atcaactgga tcatagtgtc
     1681 gtccaccaag ttcacatagt tctcataaag gtggaaagtt gaagatttga acctaagtgt
     1741 tgggtgcacc cttgacactc ccaacctgtt ccatgcgtcg aaagaagtca cgtgtctctg
     1801 caaccacaac ttctccctgt actcgttagc gtgagtaccc aatttttcaa ggaacacgta
     1861 gtttggatgt tccttcaatt tgttaccaga caatcccttt ttcattccca acttctctgt
     1921 agcgaagttg tatgacttaa ggtcagctgc ccactccttc aaattgttct tcacgtagtt
     1981 tttgtagtat ttgtaattag ggtgtgaaag aagagcgttt ccagtcaatt tatcaagctt
     2041 caacttcacc atcacgtctg ccaatggtgt gtccttctgc atccatgacc tcatcaatcc
     2101 agccacgaac tcctcgaaga acttgaaatt ctcagcctgt ggcctgtcct tcaactgaac
     2161 tgtgttaagt tcaagcctca ccaagatgtt gtcgatgtca aggtcgttct tagcccacaa
     2221 caaagcctgc tgctccatga actgagaata gtacttaaag ttagggttag aagtcaaagc
     2281 agctccctgc aaaccgtaaa tacccaactc tgtagccacc tggttaggag tccactcgtt
     2341 cttcttccac actgggatct gtgagataac gaagtcatca taaaacctaa agttagtagc
     2401 tccagtcaaa gcagatccct ccaagttgtc caatccaagt gcgttcctca cgaaagacac
     2461 tgacttcttg ttcttcctcc acacgtcaac ctttgctgaa gttgccaaaa gtggtgtcac
     2521 gtgctccctg aaagacttag ttgccttagt catagaccta gaaaatgact tcataggcct
     2581 gaatgacttc aacttctcaa aagccttaag agactgcaac ttctccaaca ctgcaccgtt
     2641 tcctgccatt gtatcgataa ttgtaaatgt aattgtaatg ttgtttgttg tttgttgttg
     2701 ttggtaattg ttgtaaaaat gagctcttat actcgagcgt gtcctctcca aatgaaatga
     2761 acttccttat atagaggaag ggtcttgcga aggatagtgg gattgtgcgt catcccttac
     2821 gtcagtggag atgtcacatc aatccacttg ctttgtagac gtggttggaa cctcttcttt
     2881 ttccacgatg ctcctcgtgg gtgggggtcc atctttggga ccactgtcgg cagagagatc
     2941 ttgaatgata gcctttcctt tatcgcaatg atggcatttg taggagccac cttccttttc
     3001 tactgtcctt tcgatgaagt gacagatagc tgggcaatgg aatccgagga ggtttcccga
     3061 aattatcctt tgttgaaaag tctcaatagc cctttgatct tctgagactg tatctttgac
     3121 atttttggag tagaccagag tgtcgtgctc caccatgttg acgaagattt tcttcttgtc
     3181 attgagtcgt aaaagactct gtatgaactg ttcgccagtc ttgacggcga gttctgttag
     3241 atcctcgatt tgaatcttag actccatgca tggccttaga ttcagtagga actacctttt
     3301 tagagactcc aatctctatt acttgccttg gtttatgaag caagccttga atcgtccata
     3361 ctggaatagt acttctgatc ttgagaaata tgtctttctc tgtgttcttg atgcaattag
     3421 tcctgaatct tttgactgca tctttaacct tcttgggaag gtatttgatc tcctggagat
     3481 tgttactcgg gtagatcgtc ttgatgagac ttgctgcgta ggcctctcta accatctgtg
     3541 ggtcagcatt ctttctgaaa ttgaagaggc taaccttctc attatcagtg gtgaacatag
     3601 tgtcgtcacc ttcaccttcg aacttccttc ctagatcgta aagatagagg aaatcgtcca
     3661 ttgtaatctc cggggcaaag gagatctctt ttggggctgg atcactgctg ggccttttgg
     3721 ttcctagcgt gagccagtgg gctttttgct ttggtgggct tgttagggcc ttagcaaagc
     3781 tcttgggctt gagttgagct tctcctttgg ggatgaagtt caacctgtct gtttgctgac
     3841 ttgttgtgta cgcgtcagct gctgctcttg cctctgtaat agtggcaaat ttcttgtgtg
     3901 caactccggg aacgccgttt gttgccgcct ttgtacaacc ccagtcatcg tatataccgg
     3961 catgtggacc gttatacaca acgtagtagt tgatatgagg gtgttgaata cccgattctg
     4021 ctctgagagg agcaactgtg ctgttaagct cagatttttg tgggattgga attcctccaa
     4081 gcttgaattc ttgcagcgct ccaattcagc aagcgtcgat ctagtaacat agatgacacc
     4141 gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc gtattaaatg
     4201 tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat gcattacatg
     4261 ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat cgcaagaccg
     4321 gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc tgcttgactc
     4381 tagatccaga gtcccgctca aatctcagtc acaggaagaa caggtctagg aggaacagga
     4441 agagagaaat caagctgcca gaatcccaca tcatgccaat ttccatgttt gaaaccagca
     4501 gctctaagca ttcccctagg agcataacca agagcctcat gcatcctcac agaaggatca
     4561 ttaggaagtc caatcacagc aaccacagac ttgaaaccct gagcctccaa agacttcaaa
     4621 agatgagtgt aaagagtaga tccaagacca gttctttgat gcctaggaga cacgtaaaca
     4681 gtagactcag cagtccaatc gtaagcattc ctagccttcc aaggaccagc ataagcaata
     4741 ccagccactt caccatccac ttcagcaaca agccaaggat acctctctct aagcctcaca
     4801 agatcatcag tccactcttg tggttcttga ggctcagtcc tgaaattcac agtagaagtc
     4861 tcaatgtaat gattcacaat agtgcacaca gcaggcatat cagcctcagt agcccttcta
     4921 atatcagcag gccttctttc aggagacatg cgaaacgatc cagatccggt gcagattatt
     4981 tggattgaga gtgaatatga gactctaatt ggataccgag gggaatttat ggaacgtcag
     5041 tggagcattt tgacaagaaa tatttgctag ctgatagtga ccttaggcga cttttgaacg
     5101 cgcaataatg gtttctgacg tatgtgctta gctcattaaa ctccagaaac ccgcggctga
     5161 gtggctcctt caacgttgcg gttctgtcag ttccaaacgt aaaacggctt gtcccgcgtc
     5221 atcggcgggg gtcataacgt gactccctta attctccgct ccggatccga attcggcaca
     5281 gagtgttcaa ccccagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt
     5341 caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca
     5401 gctcggcaca aaatcaccac tcgatacagg cagcccatca accatgacta ctgtccggtc
     5461 caacagatcc gagcaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5521 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5581 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5641 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5701 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5761 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5821 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5881 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5941 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6001 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6061 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6121 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6181 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6241 tatatgagta aacttggtct gacagctaaa acaattcatc cagtaaaata taatatttta
     6301 ttttctccca atcaggcttg atccccagta agtcaaaaaa tagctcgaca tactgttctt
     6361 ccccgatatc ctccctgagc gaccggacgc agaaggcaat gtcataccac ttgtccgccc
     6421 tgccgcttct cccaagatca ataaagccac ttactttgcc atctttcaca aagatgttgc
     6481 tgtctcccag gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa
     6541 aatcatacag ctcgcgcgga tctttaaatg gagtgtcctc ttcccagttt tcgcaatcca
     6601 catcggccag atcgttattc agtaagtaat ccaattcggc taagcggctg tctaagctat
     6661 tcgtataggg acaatccgat atgtcgatgg agtgaaagag cctgatgcac tccgcataca
     6721 gctcgataat cttttcaggg ctttgttcat cttcatactc ttccgagcaa aggacgccat
     6781 cggcctcact catgagcaga ttgctccagc catcatgccg ttcaaagtgc aggacctttg
     6841 gaacaggcag ctttccttcc agccatagca tcatgtcctt ttcccgttcc acatcatagg
     6901 tggtcccttt ataccggctg tccgtcattt ttaaatatag gttttcattt tctcccacca
     6961 gcttatatac cttagcagga gacattcctt ccgtatcttt tacgcagcgg tatttttcga
     7021 tcagtttttt caattccggt gatattctca ttttagccat actcttcctt tttcaatatt
     7081 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7141 aaaataaaca aataggggtt ccgcgcactc ggagccttga tgtgggcgct ttttgcagct
     7201 gttcggctgt gcgctggcca gacagttatg cacaggccag gcgggtttta agagttttaa
     7261 taagttttaa agagttttag gcggaaaaat cgcctttttt ctcttttata tcagtcactt
     7321 acatgtgtga ccggttccca atgtacggct ttgggttccc aatgtacggg ttccggttcc
     7381 caatgtacgg ctttgggttc ccaatgtacg tgctatccac aggaaagaag ctttttcgac
     7441 ctttttcccc tgctagggca atttgcccta gcatctgctc cgtacattag gaaccggcgg
     7501 atgcttcgcc ctcgatcagg ttgcggtagc gcatgactag gatcgggcca gcctgccccg
     7561 cctcctcctt caaatcgtac tccggcaggt catttgaccc gatcagcttg cgcacggtga
     7621 aacagaactt cttgaactct ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc
     7681 atctggcttc tgccttgcct gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc
     7741 cgggatcgat caaaaagtaa tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg
     7801 tgatctcgcg gtacatccaa tcagctagct cgatctcgat gtactccggc cgcccggttt
     7861 cgctctttac gatcttgtag cggctaatca aggcttcacc ctcggatacc gtcaccaggc
     7921 ggccgttctt ggccttcttc gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc
     7981 aggtttctac caggtcgtct ttctgctttc cgccatcggc tcgccggcag aacttgagta
     8041 cgtccgcaac gtgtggacgg aacacgcggc cgggcttgtc tcccttccct tcccggtatc
     8101 ggttcatgga ttcggttaga tgggaaaccg ccatcagtac caggtcgtaa tcccacacac
     8161 tggccatgcc ggccggccct gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga
     8221 tcacctcgcc agctcgtcgg tcacgcttcg acagacggaa aacggccacg tccatgatgc
     8281 tgcgactatc gcgggtgccc acgtcataga gcatcggaac gaaaaaatct ggttgctcgt
     8341 cgcccttggg cggcttccta atcgacgacg caccggctgc cggcggttgc cgggattctt
     8401 tgcggattcg atcagcggcc gcttgccacg attcaccggg gcgtgcttct gcctcgatgc
     8461 gttgccgctg ggcggcctgc gcggccttca acttctccac caggtcatca cccagcgccg
     8521 cgccgatttg taccgggccg gatggtttgc gaccgctcac gccgattcct cgggcttggg
     8581 ggttccagtg ccattgcagg gccggcagac aacccagccg cttacgcctg gccaaccgcc
     8641 cgttcctcca cacatggggc attccacggc gtcggtgcct ggttgttctt gattttccat
     8701 gccgcctcct ttagccgcta aaattcatct actcatttat tcatttgctc atttactctg
     8761 gtagctgcgc gatgtattca gatagcagct cggtaatggt cttgccttgg cgtaccgcgt
     8821 acatcttcag cttggtgtga tcctccgccg gcaactgaaa gttgacccgc ttcatggctg
     8881 gcgtgtctgc caggctggcc aacgttgcag ccttgctgct gcgtgcgctc ggacggccgg
     8941 cacttagcgt gtttgtgctt ttgctcattt tctctttacc tcattaactc aaatgagttt
     9001 tgatttaatt tcagcggcca gcgcctggac ctcgcgggca gcgtcgccct cgggttctga
     9061 ttcaagaacg gttgtgccgg cggcggcagt gcctgggtag ctcacgcgct gcgtgatacg
     9121 ggactcaaga atgggcagct cgtacccggc cagcgcctcg gcaacctcac cgccgatgcg
     9181 cgtgcctttg atcgcccgcg acacgacaaa ggccgcttgt agccttccat ccgtgacctc
     9241 aatgcgctgc ttaaccagct ccaccaggtc ggcggtggcc
//
