LOCUS       sequence-342195-        8302 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 8302)
  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 8302)
  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_06099
FEATURES             Location/Qualifiers
     source          1..8302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(553..735)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(589..767)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    930..954
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(1090..1678)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1170..1189)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1849..2643)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(2644..2748)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2810..3004
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(3070..4143)
                     /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(4572..5201)
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     misc_feature    5510..5534
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(5601..6308)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(6625..6970)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(6662..6682)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(8003..8255)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 gaacaggtct aggaggaaca ggaagagaga aatcaagctg ccagaatccc acatcatgcc
       61 aatttccatg tttgaaacca gcagctctaa gcattcccct aggagcataa ccaagagcct
      121 catgcatcct cacagaagga tcattaggaa gtccaatcac agcaaccaca gacttgaaac
      181 cctgagcctc caaagacttc aaaagatgag tgtaaagagt agatccaaga ccagttcttt
      241 gatgcctagg agacacgtaa acagtagact cagcagtcca atcgtaagca ttcctagcct
      301 tccaaggacc agcataagca ataccagcca cttcaccatc cacttcagca acaagccaag
      361 gatacctctc tctaagcctc acaagatcat cagtccactc ttgtggttct tgaggctcag
      421 tcctgaaatt cacagtagaa gtctcaatgt aatgattcac aatagtgcac acagcaggca
      481 tatcagcctc agtagccctt ctaatatcag caggccttct ttcaggagac atgcgaaacg
      541 atccagatcc ggtgcagatt atttggattg agagtgaata tgagactcta attggatacc
      601 gaggggaatt tatggaacgt cagtggagca ttttgacaag aaatatttgc tagctgatag
      661 tgaccttagg cgacttttga acgcgcaata atggtttctg acgtatgtgc ttagctcatt
      721 aaactccaga aacccgcggc tgagtggctc cttcaacgtt gcggttctgt cagttccaaa
      781 cgtaaaacgg cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc ttaattctcc
      841 gctccggatc cgaattcggc acagagtgtt caaccccagt acattaaaaa cgtccgcaat
      901 gtgttattaa gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc
      961 agccaacagc tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca
     1021 tcaaccatga ctactgtccg gtccaacaga tccgagcagg aaccgtaaaa aggccgcgtt
     1081 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1141 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1201 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1261 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1321 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     1381 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     1441 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     1501 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     1561 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     1621 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     1681 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     1741 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     1801 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagct aaaacaattc
     1861 atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa
     1921 aaatagctcg acatactgtt cttccccgat atcctccctg agcgaccgga cgcagaaggc
     1981 aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt
     2041 gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc
     2101 ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc
     2161 ctcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc
     2221 ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa
     2281 gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata
     2341 ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg
     2401 ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc
     2461 cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata
     2521 taggttttca ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc
     2581 ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc
     2641 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     2701 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca ctcggagcct
     2761 tgatgtgggc gctttttgca gctgttcggc tgtgcgctgg ccagacagtt atgcacaggc
     2821 caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt
     2881 tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt
     2941 cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc
     3001 cacaggaaag aagctttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg
     3061 ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac
     3121 taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga
     3181 cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg
     3241 ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag
     3301 gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag
     3361 cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc
     3421 gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc
     3481 accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac
     3541 gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc
     3601 ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt
     3661 gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag
     3721 taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg
     3781 cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg
     3841 gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg
     3901 aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg acgcaccggc
     3961 tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc
     4021 ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc
     4081 caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct
     4141 cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca gacaacccag
     4201 ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg
     4261 cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca tctactcatt
     4321 tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca gctcggtaat
     4381 ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg ccggcaactg
     4441 aaagttgacc cgcttcatgg ctggcgtgtc tgccaggctg gccaacgttg cagccttgct
     4501 gctgcgtgcg ctcggacggc cggcacttag cgtgtttgtg cttttgctca ttttctcttt
     4561 acctcattaa ctcaaatgag ttttgattta atttcagcgg ccagcgcctg gacctcgcgg
     4621 gcagcgtcgc cctcgggttc tgattcaaga acggttgtgc cggcggcggc agtgcctggg
     4681 tagctcacgc gctgcgtgat acgggactca agaatgggca gctcgtaccc ggccagcgcc
     4741 tcggcaacct caccgccgat gcgcgtgcct ttgatcgccc gcgacacgac aaaggccgct
     4801 tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag gtcggcggtg
     4861 gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc tgccttgatc
     4921 gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg
     4981 ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt
     5041 tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga atcgactaac
     5101 agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg
     5161 cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc ttgcgtattt
     5221 gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg ttttactcaa
     5281 atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag ctggccggcc
     5341 aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc acggctgact
     5401 cccgagggga accctgtggt tggcatgcac atacaaatgg acgaacggat aaaccttttc
     5461 acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg tttacccgcc
     5521 aatatatcct gtcaaacact gatagtttaa acggatcact tcgtgtccct cggtcacatg
     5581 tgcatcctct agtagcggtc ctgctgagcc tcgacatgtt gtcgcaaaat tcgccctgga
     5641 cccgcccaac gatttgtcgt cactgtcaag gtttgacctg cacttcattt ggggcccaca
     5701 tacaccaaaa aaatgctgca taattctcgg ggcagcaagt cggttacccg gccgccgtgc
     5761 tggaccgggt tgaatggtgc ccgtaacttt cggtagagcg gacggccaat actcaacttc
     5821 aaggaatctc acccatgcgc gccggcgggg aaccggagtt cccttcagtg agcgttatta
     5881 gttcgccgct cggtgtgtcg tagatactag cccctggggc acttttgaaa tttgaataag
     5941 atttatgtaa tcagtctttt aggtttgacc ggttctgccg ctttttttaa aattggattt
     6001 gtaataataa aacgcaattg tttgttattg tggcgctcta tcatagatgt cgctataaac
     6061 ctattcagca caatatattg ttttcatttt aatattgtac atataagtag tagggtacaa
     6121 tcagtaaatt gaacggagaa tattattcat aaaaatacga tagtaacggg tgatatattc
     6181 attagaatga accgaaaccg gcggtaagga tctgagctac acatgctcag gttttttaca
     6241 acgtgcacaa cagaattgaa agcaaatatc atgcgatcat aggcttctcg catatctcat
     6301 taaagcagga caagctcaga agtcctcagc cttgcagaac tcctccttcc tgttctcgta
     6361 gtaagagcag tggtcctcgt agtagtccac gtatccagtg tacacagacc tcttgatagg
     6421 gttgatagtg tcagcagtca actccaagtc cctcctcacg tccaaaggag tcatgtggtg
     6481 ctggaaccac cagtagtacc taggctggtt agaagtctgg atcaaagaga acattgtatc
     6541 gataattgta aatgtaattg taatgttgtt tgttgtttgt tgttgttggt aattgttgta
     6601 aaaatgagct cttatactcg agcgtgtcct ctccaaatga aatgaacttc cttatataga
     6661 ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatgtc
     6721 acatcaatcc acttgctttg tagacgtggt tggaacctct tctttttcca cgatgctcct
     6781 cgtgggtggg ggtccatctt tgggaccact gtcggcagag agatcttgaa tgatagcctt
     6841 tcctttatcg caatgatggc atttgtagga gccaccttcc ttttctactg tcctttcgat
     6901 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgaaatta tcctttgttg
     6961 aaaagtctca atagcccttt gatcttctga gactgtatct ttgacatttt tggagtagac
     7021 cagagtgtcg tgctccacca tgttgacgaa gattttcttc ttgtcattga gtcgtaaaag
     7081 actctgtatg aactgttcgc cagtcttgac ggcgagttct gttagatcct cgatttgaat
     7141 cttagactcc atgcatggcc ttagattcag taggaactac ctttttagag actccaatct
     7201 ctattacttg ccttggttta tgaagcaagc cttgaatcgt ccatactgga atagtacttc
     7261 tgatcttgag aaatatgtct ttctctgtgt tcttgatgca attagtcctg aatcttttga
     7321 ctgcatcttt aaccttcttg ggaaggtatt tgatctcctg gagattgtta ctcgggtaga
     7381 tcgtcttgat gagacttgct gcgtaggcct ctctaaccat ctgtgggtca gcattctttc
     7441 tgaaattgaa gaggctaacc ttctcattat cagtggtgaa catagtgtcg tcaccttcac
     7501 cttcgaactt ccttcctaga tcgtaaagat agaggaaatc gtccattgta atctccgggg
     7561 caaaggagat ctcttttggg gctggatcac tgctgggcct tttggttcct agcgtgagcc
     7621 agtgggcttt ttgctttggt gggcttgtta gggccttagc aaagctcttg ggcttgagtt
     7681 gagcttctcc tttggggatg aagttcaacc tgtctgtttg ctgacttgtt gtgtacgcgt
     7741 cagctgctgc tcttgcctct gtaatagtgg caaatttctt gtgtgcaact ccgggaacgc
     7801 cgtttgttgc cgcctttgta caaccccagt catcgtatat accggcatgt ggaccgttat
     7861 acacaacgta gtagttgata tgagggtgtt gaatacccga ttctgctctg agaggagcaa
     7921 ctgtgctgtt aagctcagat ttttgtggga ttggaattcc tccaagcttg aattcttgca
     7981 gcgctccaat tcagcaagcg tcgatctagt aacatagatg acaccgcgcg cgataattta
     8041 tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa ttgcgggact
     8101 ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat tattacatgc
     8161 ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac aggattcaat
     8221 cttaagaaac tttattgcca aatgtttgaa cgatctgctt gactctagat ccagagtccc
     8281 gctcaaatct cagtcacagg aa
//
