LOCUS       sequence-342188-        9994 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 9994)
  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 9994)
  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_04269a
FEATURES             Location/Qualifiers
     source          1..9994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    754..778
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(845..1552)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3561..3906)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3598..3618)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4939..5191)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5791..5973)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5827..6005)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    6168..6192
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6328..6916)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6408..6427)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(7087..7881)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7882..7986)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8048..8242
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8308..9381)
                     /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(9810..9994,1..445))
                     /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 cgatgcgcgt gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg
       61 tgacctcaat gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag
      121 ggcttggctg caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt
      181 ccgccgcctg gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt
      241 cgcggtcaat cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg
      301 ccgcccaatc gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg
      361 cgagttgcag ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct
      421 ggttaagtac agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg
      481 catcatatac gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt
      541 tagacggcgg cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc
      601 atcagacaaa ccggccagga tttcatgcag ccgcacggct gactcccgag gggaaccctg
      661 tggttggcat gcacatacaa atggacgaac ggataaacct tttcacgccc ttttaaatat
      721 ccgattattc taataaacgc tcttttctct taggtttacc cgccaatata tcctgtcaaa
      781 cactgatagt ttaaacggat cacttcgtgt ccctcggtca catgtgcatc ctctagtagc
      841 ggtcctgctg agcctcgaca tgttgtcgca aaattcgccc tggacccgcc caacgatttg
      901 tcgtcactgt caaggtttga cctgcacttc atttggggcc cacatacacc aaaaaaatgc
      961 tgcataattc tcggggcagc aagtcggtta cccggccgcc gtgctggacc gggttgaatg
     1021 gtgcccgtaa ctttcggtag agcggacggc caatactcaa cttcaaggaa tctcacccat
     1081 gcgcgccggc ggggaaccgg agttcccttc agtgagcgtt attagttcgc cgctcggtgt
     1141 gtcgtagata ctagcccctg gggcactttt gaaatttgaa taagatttat gtaatcagtc
     1201 ttttaggttt gaccggttct gccgcttttt ttaaaattgg atttgtaata ataaaacgca
     1261 attgtttgtt attgtggcgc tctatcatag atgtcgctat aaacctattc agcacaatat
     1321 attgttttca ttttaatatt gtacatataa gtagtagggt acaatcagta aattgaacgg
     1381 agaatattat tcataaaaat acgatagtaa cgggtgatat attcattaga atgaaccgaa
     1441 accggcggta aggatctgag ctacacatgc tcaggttttt tacaacgtgc acaacagaat
     1501 tgaaagcaaa tatcatgcga tcataggctt ctcgcatatc tcattaaagc aggacaagct
     1561 caagttggtg ttgcaaggtt ctttctaata ccaatagact taccgaatcc atttgaagag
     1621 tcgtcctggt tcattttttt tccttgcaat ttgttaaagt aagacaccca tgttccgaaa
     1681 gctgggtctg aaagtgcgtt gtctccctta ccagacaacc tcaaaagctt aaaagcctcg
     1741 tgtgcaggca ttccctctct caaccacaaa gcagatgcca cctgtgcagc agttgcctta
     1801 aggttaccag tgattgatga gtgtgtaacc atttttttca atgtctcgtc atcgtaatat
     1861 tttgtcatca ctgatgacat cacgttgtct gcctcctctt tgtgcataga cttcacgtaa
     1921 gtgaaccatg tcacaaggat aggattttcg aagaaattga taccatcctt gtgcaaagaa
     1981 aggtcagtga acacgtcctc agctgtcttt ttctgaccaa gccagttacc tttgatcaac
     2041 ctcaccatag gacccaaaga aaacttaagt gttccgtcct tcctacctgc gaagaacatc
     2101 ttaaggatct cagcgtctga gaacctgatc tcaagcttcc tgtaaagaac ctcagtaggg
     2161 ttttcagaga tctttttaac atatgagaac caagacacca atcctggaga cttcaatgtg
     2221 tcaattgatt ggtcaatgtc aagcctcaac atcttgaaaa catcgtcaga tgtttttcct
     2281 ttctccttcc agatcttgaa ctgtgtagac tccaatttaa cagccataga aaccaagtca
     2341 gagtcagact ttgccctcac aagcatctta ggcaactcgt cctctccgtg cctctttgcc
     2401 aacttcaaca ccaaaaactc atatgggtca agagtatgat gtttagtagc gtaagagata
     2461 aaagtttgaa gagcaggatg tttcaacaaa cccaactcct tgtctgcagg cacaagagag
     2521 atcttaagca cgtcgtcctc tgccttacct gtagtcaacc acttcttgat ctgtgcgaac
     2581 tccaaacctg aagcgatgtc cctagtagaa gctgtagttg atgctgccat aagcctcttg
     2641 tgcaatgcgt cctcctcata ctgcttagca agcttcaaca aaagaaagtc atatgggtcc
     2701 tccttcaact tagtcttaac gaatgaaatc catgttggaa gaagagggtt cacaagcacg
     2761 tctgcatcct ccacgttaag tttaagcaag ttataaatat ctcctgcagt ctttttctgt
     2821 gtcacccagt tgtcgaattg gatagccttc accattgtag ccaccttaga tgctcctggc
     2881 ctctgagcat ccacaaggat ctttgtaaga gcctccacgt cgtacttagc tgtaagagca
     2941 gaatggaaag cctcctcgtt aagctcaggc cttttgttat tgatagtagt cacagactgt
     3001 gcaagtgctg tgtcagtcct ctttgactcc attgcaccca ctccaagctt agcaacagag
     3061 tcctctgttc ccttctggtt gttgaaagcc aatctatttt tcaccctaga agcaaacttt
     3121 tccaattttg tagtaccaga agtgatcaat cccttcaact tctcagctcc tggcaccttg
     3181 atgtccactc tctcctcaga tgtatcgtac tcatcatcct ttgctgacca tgttttagta
     3241 cccttgtctg ggcatccgaa tgtcctgcaa ggcctagcct cagaagcctc gaaagaaccc
     3301 acagtagaat caatgaaacc gatgcacctg tcgataggtg aacctctagc aagcaacttt
     3361 gagcacttag cgtagataag gtcaaccata aacctgaaga taaggcaaag tgctgatgga
     3421 gatctaccga acatcacagc caagtcaccg tacctaccag gataagccat tgtatcgata
     3481 attgtaaatg taattgtaat gttgtttgtt gtttgttgtt gttggtaatt gttgtaaaaa
     3541 tgagctctta tactcgagcg tgtcctctcc aaatgaaatg aacttcctta tatagaggaa
     3601 gggtcttgcg aaggatagtg ggattgtgcg tcatccctta cgtcagtgga gatgtcacat
     3661 caatccactt gctttgtaga cgtggttgga acctcttctt tttccacgat gctcctcgtg
     3721 ggtgggggtc catctttggg accactgtcg gcagagagat cttgaatgat agcctttcct
     3781 ttatcgcaat gatggcattt gtaggagcca ccttcctttt ctactgtcct ttcgatgaag
     3841 tgacagatag ctgggcaatg gaatccgagg aggtttcccg aaattatcct ttgttgaaaa
     3901 gtctcaatag ccctttgatc ttctgagact gtatctttga catttttgga gtagaccaga
     3961 gtgtcgtgct ccaccatgtt gacgaagatt ttcttcttgt cattgagtcg taaaagactc
     4021 tgtatgaact gttcgccagt cttgacggcg agttctgtta gatcctcgat ttgaatctta
     4081 gactccatgc atggccttag attcagtagg aactaccttt ttagagactc caatctctat
     4141 tacttgcctt ggtttatgaa gcaagccttg aatcgtccat actggaatag tacttctgat
     4201 cttgagaaat atgtctttct ctgtgttctt gatgcaatta gtcctgaatc ttttgactgc
     4261 atctttaacc ttcttgggaa ggtatttgat ctcctggaga ttgttactcg ggtagatcgt
     4321 cttgatgaga cttgctgcgt aggcctctct aaccatctgt gggtcagcat tctttctgaa
     4381 attgaagagg ctaaccttct cattatcagt ggtgaacata gtgtcgtcac cttcaccttc
     4441 gaacttcctt cctagatcgt aaagatagag gaaatcgtcc attgtaatct ccggggcaaa
     4501 ggagatctct tttggggctg gatcactgct gggccttttg gttcctagcg tgagccagtg
     4561 ggctttttgc tttggtgggc ttgttagggc cttagcaaag ctcttgggct tgagttgagc
     4621 ttctcctttg gggatgaagt tcaacctgtc tgtttgctga cttgttgtgt acgcgtcagc
     4681 tgctgctctt gcctctgtaa tagtggcaaa tttcttgtgt gcaactccgg gaacgccgtt
     4741 tgttgccgcc tttgtacaac cccagtcatc gtatataccg gcatgtggac cgttatacac
     4801 aacgtagtag ttgatatgag ggtgttgaat acccgattct gctctgagag gagcaactgt
     4861 gctgttaagc tcagattttt gtgggattgg aattcctcca agcttgaatt cttgcagcgc
     4921 tccaattcag caagcgtcga tctagtaaca tagatgacac cgcgcgcgat aatttatcct
     4981 agtttgcgcg ctatattttg ttttctatcg cgtattaaat gtataattgc gggactctaa
     5041 tcataaaaac ccatctcata aataacgtca tgcattacat gttaattatt acatgcttaa
     5101 cgtaattcaa cagaaattat atgataatca tcgcaagacc ggcaacagga ttcaatctta
     5161 agaaacttta ttgccaaatg tttgaacgat ctgcttgact ctagatccag agtcccgctc
     5221 aaatctcagt cacaggaaga acaggtctag gaggaacagg aagagagaaa tcaagctgcc
     5281 agaatcccac atcatgccaa tttccatgtt tgaaaccagc agctctaagc attcccctag
     5341 gagcataacc aagagcctca tgcatcctca cagaaggatc attaggaagt ccaatcacag
     5401 caaccacaga cttgaaaccc tgagcctcca aagacttcaa aagatgagtg taaagagtag
     5461 atccaagacc agttctttga tgcctaggag acacgtaaac agtagactca gcagtccaat
     5521 cgtaagcatt cctagccttc caaggaccag cataagcaat accagccact tcaccatcca
     5581 cttcagcaac aagccaagga tacctctctc taagcctcac aagatcatca gtccactctt
     5641 gtggttcttg aggctcagtc ctgaaattca cagtagaagt ctcaatgtaa tgattcacaa
     5701 tagtgcacac agcaggcata tcagcctcag tagcccttct aatatcagca ggccttcttt
     5761 caggagacat gcgaaacgat ccagatccgg tgcagattat ttggattgag agtgaatatg
     5821 agactctaat tggataccga ggggaattta tggaacgtca gtggagcatt ttgacaagaa
     5881 atatttgcta gctgatagtg accttaggcg acttttgaac gcgcaataat ggtttctgac
     5941 gtatgtgctt agctcattaa actccagaaa cccgcggctg agtggctcct tcaacgttgc
     6001 ggttctgtca gttccaaacg taaaacggct tgtcccgcgt catcggcggg ggtcataacg
     6061 tgactccctt aattctccgc tccggatccg aattcggcac agagtgttca accccagtac
     6121 attaaaaacg tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa
     6181 tatatcctgc caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca
     6241 ctcgatacag gcagcccatc aaccatgact actgtccggt ccaacagatc cgagcaggaa
     6301 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6361 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6421 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6481 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6541 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6601 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6661 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     6721 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     6781 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     6841 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     6901 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     6961 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     7021 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     7081 tgacagctaa aacaattcat ccagtaaaat ataatatttt attttctccc aatcaggctt
     7141 gatccccagt aagtcaaaaa atagctcgac atactgttct tccccgatat cctccctgag
     7201 cgaccggacg cagaaggcaa tgtcatacca cttgtccgcc ctgccgcttc tcccaagatc
     7261 aataaagcca cttactttgc catctttcac aaagatgttg ctgtctccca ggtcgccgtg
     7321 ggaaaagaca agttcctctt cgggcttttc cgtctttaaa aaatcataca gctcgcgcgg
     7381 atctttaaat ggagtgtcct cttcccagtt ttcgcaatcc acatcggcca gatcgttatt
     7441 cagtaagtaa tccaattcgg ctaagcggct gtctaagcta ttcgtatagg gacaatccga
     7501 tatgtcgatg gagtgaaaga gcctgatgca ctccgcatac agctcgataa tcttttcagg
     7561 gctttgttca tcttcatact cttccgagca aaggacgcca tcggcctcac tcatgagcag
     7621 attgctccag ccatcatgcc gttcaaagtg caggaccttt ggaacaggca gctttccttc
     7681 cagccatagc atcatgtcct tttcccgttc cacatcatag gtggtccctt tataccggct
     7741 gtccgtcatt tttaaatata ggttttcatt ttctcccacc agcttatata ccttagcagg
     7801 agacattcct tccgtatctt ttacgcagcg gtatttttcg atcagttttt tcaattccgg
     7861 tgatattctc attttagcca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7921 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7981 tccgcgcact cggagccttg atgtgggcgc tttttgcagc tgttcggctg tgcgctggcc
     8041 agacagttat gcacaggcca ggcgggtttt aagagtttta ataagtttta aagagtttta
     8101 ggcggaaaaa tcgccttttt tctcttttat atcagtcact tacatgtgtg accggttccc
     8161 aatgtacggc tttgggttcc caatgtacgg gttccggttc ccaatgtacg gctttgggtt
     8221 cccaatgtac gtgctatcca caggaaagaa gctttttcga cctttttccc ctgctagggc
     8281 aatttgccct agcatctgct ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag
     8341 gttgcggtag cgcatgacta ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta
     8401 ctccggcagg tcatttgacc cgatcagctt gcgcacggtg aaacagaact tcttgaactc
     8461 tccggcgctg ccactgcgtt cgtagatcgt cttgaacaac catctggctt ctgccttgcc
     8521 tgcggcgcgg cgtgccaggc ggtagagaaa acggccgatg ccgggatcga tcaaaaagta
     8581 atcggggtga accgtcagca cgtccgggtt cttgccttct gtgatctcgc ggtacatcca
     8641 atcagctagc tcgatctcga tgtactccgg ccgcccggtt tcgctcttta cgatcttgta
     8701 gcggctaatc aaggcttcac cctcggatac cgtcaccagg cggccgttct tggccttctt
     8761 cgtacgctgc atggcaacgt gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc
     8821 tttctgcttt ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg
     8881 gaacacgcgg ccgggcttgt ctcccttccc ttcccggtat cggttcatgg attcggttag
     8941 atgggaaacc gccatcagta ccaggtcgta atcccacaca ctggccatgc cggccggccc
     9001 tgcggaaacc tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg
     9061 gtcacgcttc gacagacgga aaacggccac gtccatgatg ctgcgactat cgcgggtgcc
     9121 cacgtcatag agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct
     9181 aatcgacgac gcaccggctg ccggcggttg ccgggattct ttgcggattc gatcagcggc
     9241 cgcttgccac gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg
     9301 cgcggccttc aacttctcca ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc
     9361 ggatggtttg cgaccgctca cgccgattcc tcgggcttgg gggttccagt gccattgcag
     9421 ggccggcaga caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg
     9481 cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct
     9541 aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc
     9601 agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg
     9661 atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc
     9721 caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct
     9781 tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc
     9841 agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg
     9901 gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc
     9961 tcgtacccgg ccagcgcctc ggcaacctca ccgc
//
