LOCUS       sequence-342327-        9916 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 9916)
  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 9916)
  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_12057a
FEATURES             Location/Qualifiers
     source          1..9916
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(226..855)
                     /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    1164..1188
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1255..1962)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3893..4238)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3930..3950)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(5271..5523)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(6123..6305)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(6159..6337)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6500..6524
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6660..7248)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6740..6759)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7419..8213)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(8214..8318)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8380..8574
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8640..9713)
                     /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"
ORIGIN
        1 gctgcgcgat gtattcagat agcagctcgg taatggtctt gccttggcgt accgcgtaca
       61 tcttcagctt ggtgtgatcc tccgccggca actgaaagtt gacccgcttc atggctggcg
      121 tgtctgccag gctggccaac gttgcagcct tgctgctgcg tgcgctcgga cggccggcac
      181 ttagcgtgtt tgtgcttttg ctcattttct ctttacctca ttaactcaaa tgagttttga
      241 tttaatttca gcggccagcg cctggacctc gcgggcagcg tcgccctcgg gttctgattc
      301 aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg tgatacggga
      361 ctcaagaatg ggcagctcgt acccggccag cgcctcggca acctcaccgc cgatgcgcgt
      421 gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg tgacctcaat
      481 gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg
      541 caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt ccgccgcctg
      601 gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat
      661 cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg ccgcccaatc
      721 gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg cgagttgcag
      781 ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct ggttaagtac
      841 agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg catcatatac
      901 gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt tagacggcgg
      961 cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc atcagacaaa
     1021 ccggccagga tttcatgcag ccgcacggct gactcccgag gggaaccctg tggttggcat
     1081 gcacatacaa atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc
     1141 taataaacgc tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt
     1201 ttaaacggat cacttcgtgt ccctcggtca catgtgcatc ctctagtagc ggtcctgctg
     1261 agcctcgaca tgttgtcgca aaattcgccc tggacccgcc caacgatttg tcgtcactgt
     1321 caaggtttga cctgcacttc atttggggcc cacatacacc aaaaaaatgc tgcataattc
     1381 tcggggcagc aagtcggtta cccggccgcc gtgctggacc gggttgaatg gtgcccgtaa
     1441 ctttcggtag agcggacggc caatactcaa cttcaaggaa tctcacccat gcgcgccggc
     1501 ggggaaccgg agttcccttc agtgagcgtt attagttcgc cgctcggtgt gtcgtagata
     1561 ctagcccctg gggcactttt gaaatttgaa taagatttat gtaatcagtc ttttaggttt
     1621 gaccggttct gccgcttttt ttaaaattgg atttgtaata ataaaacgca attgtttgtt
     1681 attgtggcgc tctatcatag atgtcgctat aaacctattc agcacaatat attgttttca
     1741 ttttaatatt gtacatataa gtagtagggt acaatcagta aattgaacgg agaatattat
     1801 tcataaaaat acgatagtaa cgggtgatat attcattaga atgaaccgaa accggcggta
     1861 aggatctgag ctacacatgc tcaggttttt tacaacgtgc acaacagaat tgaaagcaaa
     1921 tatcatgcga tcataggctt ctcgcatatc tcattaaagc aggacaagct caatttttct
     1981 tgtacaaaga caaagcgtgc tccacgtctg atgccaactt ctctgttgta gggtttgact
     2041 tagcagtctt aatcatctga aacaattcat cctcgtcgta gtacttcaag aaagtagtaa
     2101 tcatatctgg tttttgctta tagtaagtag ccatgtattg gatccagatc tccaagaatg
     2161 gtgaagcaaa aagagtctta cctgccaact caaggtccaa tccctcgaag atctcgtctg
     2221 gtgtcctagt ctttcttgac acgttccagt ccttaagaag ctccctttcc atcctctttg
     2281 caattttttc agttgaagaa actttcatac ctgaaatgat cattttcaca agtgacttat
     2341 cgtcgtggtg cctcctcact gttttcatca tagtcctcct caagtctgga aatgtcaaat
     2401 aaaagtcgtc aaggtactga atccatgttg cgaactgtgg agaagccaac aagtcgtcac
     2461 ccttctctgc caacctaagc attgagaatg tgtcagaagg caactgaccc accatcctct
     2521 ccacaagctc gatattaact ccagtagcag caagcatagt ccagatagta gtctccttac
     2581 cgtggttctc cctgttgtaa gcagacaaat acttacccca tgtgatcacc aatggattag
     2641 tgaaaagctc atctttttcc tgtgcaaggt ggaaaacatt attgaaaagg tattcaggag
     2701 tccatccctt ctgcatccac ctctggaaaa gctctgactg aagcttctta gctgcacctg
     2761 cagtgtcctt cacatgtgtt gccttatcca acaatgaaac aaggttaagc tctccgtagt
     2821 ttgctgccat tgtaccaata agagttgcct tgtactttgg gtttgcagcg ttatactcct
     2881 tcatgtaatt aatccaagtc ttaagctgag ggtttgaaag cacgtcgtca tcgaacttgt
     2941 ccaacttcaa ctgcttgaag aatgcgtcag gagttccctt agccaaccac ttctgctcag
     3001 tctcaagttc cctaataagc tttgcagatc ctggcacgtt cttagcagac tcaagcatct
     3061 tgataagtcc gtcagagtcg tagtgtgctg ccaaaacaga catcaaagat gttcttttat
     3121 cagggttagc agtattcaaa tcatccacgt aagaaaccca cttcataaac tgagggtggt
     3181 cgaaaagctt ctcacctgct ttagacaaag acaagaactg aaacacatcc ctaggtgact
     3241 tctctgtgta aagccatctt ttgatctgct cgtcctgcaa gtttgctgca agcttagctg
     3301 ttgaaggaga cttcattgcg tcgaaaagca tagcagtaag cttgtcatct ccgaagtaat
     3361 ttgtcataac agcaatccta gagattctag tatttttaga gttgaaatcg tcaaggtatg
     3421 taagccaagt tgcgaactga gggttctcca aaagcttgtc tcctgccttg tccaatgcca
     3481 aaaacttgaa cacgtccctt ggcctctttc cagcgtcaag ccacaattga atttgttcat
     3541 tgtgcaattt accagccaca ttagctgtag aaggtgtttt catagccctg atcaagattg
     3601 ttgaaagtgc ctcgtcaccg tagtgtgtag tcaagatagg gatagactca tccacctttg
     3661 acattgtttt ctctttaaag tttgttgcgt aagaaagcca agctgttgca agtgggttct
     3721 ccaacaattt atcacctgct tttgaaagtt ccaaaagctt aaacacgtca gctggagctt
     3781 tcttgttgtc caaccagtac attgtatcga taattgtaaa tgtaattgta atgttgtttg
     3841 ttgtttgttg ttgttggtaa ttgttgtaaa aatgagctct tatactcgag cgtgtcctct
     3901 ccaaatgaaa tgaacttcct tatatagagg aagggtcttg cgaaggatag tgggattgtg
     3961 cgtcatccct tacgtcagtg gagatgtcac atcaatccac ttgctttgta gacgtggttg
     4021 gaacctcttc tttttccacg atgctcctcg tgggtggggg tccatctttg ggaccactgt
     4081 cggcagagag atcttgaatg atagcctttc ctttatcgca atgatggcat ttgtaggagc
     4141 caccttcctt ttctactgtc ctttcgatga agtgacagat agctgggcaa tggaatccga
     4201 ggaggtttcc cgaaattatc ctttgttgaa aagtctcaat agccctttga tcttctgaga
     4261 ctgtatcttt gacatttttg gagtagacca gagtgtcgtg ctccaccatg ttgacgaaga
     4321 ttttcttctt gtcattgagt cgtaaaagac tctgtatgaa ctgttcgcca gtcttgacgg
     4381 cgagttctgt tagatcctcg atttgaatct tagactccat gcatggcctt agattcagta
     4441 ggaactacct ttttagagac tccaatctct attacttgcc ttggtttatg aagcaagcct
     4501 tgaatcgtcc atactggaat agtacttctg atcttgagaa atatgtcttt ctctgtgttc
     4561 ttgatgcaat tagtcctgaa tcttttgact gcatctttaa ccttcttggg aaggtatttg
     4621 atctcctgga gattgttact cgggtagatc gtcttgatga gacttgctgc gtaggcctct
     4681 ctaaccatct gtgggtcagc attctttctg aaattgaaga ggctaacctt ctcattatca
     4741 gtggtgaaca tagtgtcgtc accttcacct tcgaacttcc ttcctagatc gtaaagatag
     4801 aggaaatcgt ccattgtaat ctccggggca aaggagatct cttttggggc tggatcactg
     4861 ctgggccttt tggttcctag cgtgagccag tgggcttttt gctttggtgg gcttgttagg
     4921 gccttagcaa agctcttggg cttgagttga gcttctcctt tggggatgaa gttcaacctg
     4981 tctgtttgct gacttgttgt gtacgcgtca gctgctgctc ttgcctctgt aatagtggca
     5041 aatttcttgt gtgcaactcc gggaacgccg tttgttgccg cctttgtaca accccagtca
     5101 tcgtatatac cggcatgtgg accgttatac acaacgtagt agttgatatg agggtgttga
     5161 atacccgatt ctgctctgag aggagcaact gtgctgttaa gctcagattt ttgtgggatt
     5221 ggaattcctc caagcttgaa ttcttgcagc gctccaattc agcaagcgtc gatctagtaa
     5281 catagatgac accgcgcgcg ataatttatc ctagtttgcg cgctatattt tgttttctat
     5341 cgcgtattaa atgtataatt gcgggactct aatcataaaa acccatctca taaataacgt
     5401 catgcattac atgttaatta ttacatgctt aacgtaattc aacagaaatt atatgataat
     5461 catcgcaaga ccggcaacag gattcaatct taagaaactt tattgccaaa tgtttgaacg
     5521 atctgcttga ctctagatcc agagtcccgc tcaaatctca gtcacaggaa gaacaggtct
     5581 aggaggaaca ggaagagaga aatcaagctg ccagaatccc acatcatgcc aatttccatg
     5641 tttgaaacca gcagctctaa gcattcccct aggagcataa ccaagagcct catgcatcct
     5701 cacagaagga tcattaggaa gtccaatcac agcaaccaca gacttgaaac cctgagcctc
     5761 caaagacttc aaaagatgag tgtaaagagt agatccaaga ccagttcttt gatgcctagg
     5821 agacacgtaa acagtagact cagcagtcca atcgtaagca ttcctagcct tccaaggacc
     5881 agcataagca ataccagcca cttcaccatc cacttcagca acaagccaag gatacctctc
     5941 tctaagcctc acaagatcat cagtccactc ttgtggttct tgaggctcag tcctgaaatt
     6001 cacagtagaa gtctcaatgt aatgattcac aatagtgcac acagcaggca tatcagcctc
     6061 agtagccctt ctaatatcag caggccttct ttcaggagac atgcgaaacg atccagatcc
     6121 ggtgcagatt atttggattg agagtgaata tgagactcta attggatacc gaggggaatt
     6181 tatggaacgt cagtggagca ttttgacaag aaatatttgc tagctgatag tgaccttagg
     6241 cgacttttga acgcgcaata atggtttctg acgtatgtgc ttagctcatt aaactccaga
     6301 aacccgcggc tgagtggctc cttcaacgtt gcggttctgt cagttccaaa cgtaaaacgg
     6361 cttgtcccgc gtcatcggcg ggggtcataa cgtgactccc ttaattctcc gctccggatc
     6421 cgaattcggc acagagtgtt caaccccagt acattaaaaa cgtccgcaat gtgttattaa
     6481 gttgtctaag cgtcaatttg tttacaccac aatatatcct gccaccagcc agccaacagc
     6541 tccccgaccg gcagctcggc acaaaatcac cactcgatac aggcagccca tcaaccatga
     6601 ctactgtccg gtccaacaga tccgagcagg aaccgtaaaa aggccgcgtt gctggcgttt
     6661 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6721 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6781 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6841 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6901 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6961 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7021 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7081 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7141 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7201 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7261 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7321 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7381 tcaatctaaa gtatatatga gtaaacttgg tctgacagct aaaacaattc atccagtaaa
     7441 atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa aaatagctcg
     7501 acatactgtt cttccccgat atcctccctg agcgaccgga cgcagaaggc aatgtcatac
     7561 cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt gccatctttc
     7621 acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc ttcgggcttt
     7681 tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc ctcttcccag
     7741 ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc ggctaagcgg
     7801 ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa gagcctgatg
     7861 cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata ctcttccgag
     7921 caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg ccgttcaaag
     7981 tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc cttttcccgt
     8041 tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata taggttttca
     8101 ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc ttttacgcag
     8161 cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc catactcttc
     8221 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     8281 gaatgtattt agaaaaataa acaaataggg gttccgcgca ctcggagcct tgatgtgggc
     8341 gctttttgca gctgttcggc tgtgcgctgg ccagacagtt atgcacaggc caggcgggtt
     8401 ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt tttctctttt
     8461 atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt cccaatgtac
     8521 gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc cacaggaaag
     8581 aagctttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg ctccgtacat
     8641 taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac taggatcggg
     8701 ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga cccgatcagc
     8761 ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg ttcgtagatc
     8821 gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag gcggtagaga
     8881 aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag cacgtccggg
     8941 ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc gatgtactcc
     9001 ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc accctcggat
     9061 accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac gtgcgtggtg
     9121 tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc ggctcgccgg
     9181 cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt gtctcccttc
     9241 ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag taccaggtcg
     9301 taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg cccgtctgga
     9361 agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg gaaaacggcc
     9421 acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg aacgaaaaaa
     9481 tctggttgct cgtcgccctt gggcggcttc ctaatcgacg acgcaccggc tgccggcggt
     9541 tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc ggggcgtgct
     9601 tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc caccaggtca
     9661 tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct cacgccgatt
     9721 cctcgggctt gggggttcca gtgccattgc agggccggca gacaacccag ccgcttacgc
     9781 ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg cctggttgtt
     9841 cttgattttc catgccgcct cctttagccg ctaaaattca tctactcatt tattcatttg
     9901 ctcatttact ctggta
//
