LOCUS       sequence-342198-        9304 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 9304)
  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 9304)
  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_06478
FEATURES             Location/Qualifiers
     source          1..9304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    683..707
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(774..1481)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     CDS             complement(1772..1783)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(2800..3145)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2837..2857)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4178..4430)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5030..5212)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5066..5244)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5407..5431
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5567..6155)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5647..5666)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6326..7120)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7121..7225)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7287..7481
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7547..8620)
                     /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(9049..9304,1..374))
                     /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 cgctgcttaa ccagctccac caggtcggcg gtggcccata tgtcgtaagg gcttggctgc
       61 accggaatca gcacgaagtc ggctgccttg atcgcggaca cagccaagtc cgccgcctgg
      121 ggcgctccgt cgatcactac gaagtcgcgc cggccgatgg ccttcacgtc gcggtcaatc
      181 gtcgggcggt cgatgccgac aacggttagc ggttgatctt cccgcacggc cgcccaatcg
      241 cgggcactgc cctggggatc ggaatcgact aacagaacat cggccccggc gagttgcagg
      301 gcgcgggcta gatgggttgc gatggtcgtc ttgcctgacc cgcctttctg gttaagtaca
      361 gcgataacct tcatgcgttc cccttgcgta tttgtttatt tactcatcgc atcatatacg
      421 cagcgaccgc atgacgcaag ctgttttact caaatacaca tcaccttttt agacggcggc
      481 gctcggtttc ttcagcggcc aagctggccg gccaggccgc cagcttggca tcagacaaac
      541 cggccaggat ttcatgcagc cgcacggctg actcccgagg ggaaccctgt ggttggcatg
      601 cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc cgattattct
      661 aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac actgatagtt
      721 taaacggatc acttcgtgtc cctcggtcac atgtgcatcc tctagtagcg gtcctgctga
      781 gcctcgacat gttgtcgcaa aattcgccct ggacccgccc aacgatttgt cgtcactgtc
      841 aaggtttgac ctgcacttca tttggggccc acatacacca aaaaaatgct gcataattct
      901 cggggcagca agtcggttac ccggccgccg tgctggaccg ggttgaatgg tgcccgtaac
      961 tttcggtaga gcggacggcc aatactcaac ttcaaggaat ctcacccatg cgcgccggcg
     1021 gggaaccgga gttcccttca gtgagcgtta ttagttcgcc gctcggtgtg tcgtagatac
     1081 tagcccctgg ggcacttttg aaatttgaat aagatttatg taatcagtct tttaggtttg
     1141 accggttctg ccgctttttt taaaattgga tttgtaataa taaaacgcaa ttgtttgtta
     1201 ttgtggcgct ctatcataga tgtcgctata aacctattca gcacaatata ttgttttcat
     1261 tttaatattg tacatataag tagtagggta caatcagtaa attgaacgga gaatattatt
     1321 cataaaaata cgatagtaac gggtgatata ttcattagaa tgaaccgaaa ccggcggtaa
     1381 ggatctgagc tacacatgct caggtttttt acaacgtgca caacagaatt gaaagcaaat
     1441 atcatgcgat cataggcttc tcgcatatct cattaaagca ggacaagctc attgtgcctt
     1501 catagctgca gtaagagctc cgattgggtt atcgttgtca agaagtgttt taaccttgtt
     1561 aagcaaagac actcctccct tcctctctgt gtagtacaat gtgtaagcaa cccaagtttt
     1621 gtaaattggg tcgttcttag gcaaatacac agcagaagta gggaatggga tagcaagcat
     1681 agatgacaca gccttagggt cccactcagt gtgcaaccaa ttctggaaca aataagtttg
     1741 catgttctca gcaagcctct taaggtcagg cacgtccttc aatgactcaa agaatgtagc
     1801 gaactgccac tctgacctca atggctttgt gtcagcaagg aagtgcacca tctgcacaga
     1861 tgacaacacg ttcttctttg tgtgagatct gtacaaatta acgtacctaa gccaatggat
     1921 gaacattgcg tttgtgtcca cagcgtccat gttcaccttc acaagcctaa ggattttgaa
     1981 cacctgctca ggtgtcacgt cgtaattcaa ccacacgtcg tgcatcatct tcctagaagt
     2041 ctttgaagaa aggaacaaga acctctgcat tgtgtcaccg tggtccttca ttccaggcac
     2101 gttcctcaaa aggtgcaaca atgtcaccac gtccttctct ggcattgttt ttctgaaaag
     2161 gtcaagcatg tcgtcgtcac cgaaccagtg gtttcccttc tgagccctgt actgctgagc
     2221 gtatgccaac cactggtgga agattggaga gtccaccaac tttgttccag cctcctgcaa
     2281 cctcaatgta tcgtacacaa tcttaggatc agtctcgtgc tgcaaagttt tagacaccca
     2341 tgaaccagac aaagattttt gcatagactt tcccaactcc ttaagagctc ctgtctttct
     2401 gaatgactcg atcaagatct tcaattcctc tcctgaagat ctcttcaaca acaagtagta
     2461 caagtcaagg tctgagaaag cctggtttcc gttctttgcc ctgaaagcag ccaccaactt
     2521 aagccacaac aacaaattag gatcgtcgtt aagctttgta gtagattttg ccaatctcaa
     2581 tgtgtcgaaa atagcctgag ggtgcaccct agacttcatc caagccttga tctgagcctg
     2641 ctcagggtta gacttgaaca ccttatggat aagatcgtca accttagaca aaagtggagt
     2701 agttcccatt gtatcgataa ttgtaaatgt aattgtaatg ttgtttgttg tttgttgttg
     2761 ttggtaattg ttgtaaaaat gagctcttat actcgagcgt gtcctctcca aatgaaatga
     2821 acttccttat atagaggaag ggtcttgcga aggatagtgg gattgtgcgt catcccttac
     2881 gtcagtggag atgtcacatc aatccacttg ctttgtagac gtggttggaa cctcttcttt
     2941 ttccacgatg ctcctcgtgg gtgggggtcc atctttggga ccactgtcgg cagagagatc
     3001 ttgaatgata gcctttcctt tatcgcaatg atggcatttg taggagccac cttccttttc
     3061 tactgtcctt tcgatgaagt gacagatagc tgggcaatgg aatccgagga ggtttcccga
     3121 aattatcctt tgttgaaaag tctcaatagc cctttgatct tctgagactg tatctttgac
     3181 atttttggag tagaccagag tgtcgtgctc caccatgttg acgaagattt tcttcttgtc
     3241 attgagtcgt aaaagactct gtatgaactg ttcgccagtc ttgacggcga gttctgttag
     3301 atcctcgatt tgaatcttag actccatgca tggccttaga ttcagtagga actacctttt
     3361 tagagactcc aatctctatt acttgccttg gtttatgaag caagccttga atcgtccata
     3421 ctggaatagt acttctgatc ttgagaaata tgtctttctc tgtgttcttg atgcaattag
     3481 tcctgaatct tttgactgca tctttaacct tcttgggaag gtatttgatc tcctggagat
     3541 tgttactcgg gtagatcgtc ttgatgagac ttgctgcgta ggcctctcta accatctgtg
     3601 ggtcagcatt ctttctgaaa ttgaagaggc taaccttctc attatcagtg gtgaacatag
     3661 tgtcgtcacc ttcaccttcg aacttccttc ctagatcgta aagatagagg aaatcgtcca
     3721 ttgtaatctc cggggcaaag gagatctctt ttggggctgg atcactgctg ggccttttgg
     3781 ttcctagcgt gagccagtgg gctttttgct ttggtgggct tgttagggcc ttagcaaagc
     3841 tcttgggctt gagttgagct tctcctttgg ggatgaagtt caacctgtct gtttgctgac
     3901 ttgttgtgta cgcgtcagct gctgctcttg cctctgtaat agtggcaaat ttcttgtgtg
     3961 caactccggg aacgccgttt gttgccgcct ttgtacaacc ccagtcatcg tatataccgg
     4021 catgtggacc gttatacaca acgtagtagt tgatatgagg gtgttgaata cccgattctg
     4081 ctctgagagg agcaactgtg ctgttaagct cagatttttg tgggattgga attcctccaa
     4141 gcttgaattc ttgcagcgct ccaattcagc aagcgtcgat ctagtaacat agatgacacc
     4201 gcgcgcgata atttatccta gtttgcgcgc tatattttgt tttctatcgc gtattaaatg
     4261 tataattgcg ggactctaat cataaaaacc catctcataa ataacgtcat gcattacatg
     4321 ttaattatta catgcttaac gtaattcaac agaaattata tgataatcat cgcaagaccg
     4381 gcaacaggat tcaatcttaa gaaactttat tgccaaatgt ttgaacgatc tgcttgactc
     4441 tagatccaga gtcccgctca aatctcagtc acaggaagaa caggtctagg aggaacagga
     4501 agagagaaat caagctgcca gaatcccaca tcatgccaat ttccatgttt gaaaccagca
     4561 gctctaagca ttcccctagg agcataacca agagcctcat gcatcctcac agaaggatca
     4621 ttaggaagtc caatcacagc aaccacagac ttgaaaccct gagcctccaa agacttcaaa
     4681 agatgagtgt aaagagtaga tccaagacca gttctttgat gcctaggaga cacgtaaaca
     4741 gtagactcag cagtccaatc gtaagcattc ctagccttcc aaggaccagc ataagcaata
     4801 ccagccactt caccatccac ttcagcaaca agccaaggat acctctctct aagcctcaca
     4861 agatcatcag tccactcttg tggttcttga ggctcagtcc tgaaattcac agtagaagtc
     4921 tcaatgtaat gattcacaat agtgcacaca gcaggcatat cagcctcagt agcccttcta
     4981 atatcagcag gccttctttc aggagacatg cgaaacgatc cagatccggt gcagattatt
     5041 tggattgaga gtgaatatga gactctaatt ggataccgag gggaatttat ggaacgtcag
     5101 tggagcattt tgacaagaaa tatttgctag ctgatagtga ccttaggcga cttttgaacg
     5161 cgcaataatg gtttctgacg tatgtgctta gctcattaaa ctccagaaac ccgcggctga
     5221 gtggctcctt caacgttgcg gttctgtcag ttccaaacgt aaaacggctt gtcccgcgtc
     5281 atcggcgggg gtcataacgt gactccctta attctccgct ccggatccga attcggcaca
     5341 gagtgttcaa ccccagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt
     5401 caatttgttt acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca
     5461 gctcggcaca aaatcaccac tcgatacagg cagcccatca accatgacta ctgtccggtc
     5521 caacagatcc gagcaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     5581 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     5641 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     5701 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     5761 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     5821 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     5881 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5941 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     6001 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     6061 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     6121 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6181 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6241 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6301 tatatgagta aacttggtct gacagctaaa acaattcatc cagtaaaata taatatttta
     6361 ttttctccca atcaggcttg atccccagta agtcaaaaaa tagctcgaca tactgttctt
     6421 ccccgatatc ctccctgagc gaccggacgc agaaggcaat gtcataccac ttgtccgccc
     6481 tgccgcttct cccaagatca ataaagccac ttactttgcc atctttcaca aagatgttgc
     6541 tgtctcccag gtcgccgtgg gaaaagacaa gttcctcttc gggcttttcc gtctttaaaa
     6601 aatcatacag ctcgcgcgga tctttaaatg gagtgtcctc ttcccagttt tcgcaatcca
     6661 catcggccag atcgttattc agtaagtaat ccaattcggc taagcggctg tctaagctat
     6721 tcgtataggg acaatccgat atgtcgatgg agtgaaagag cctgatgcac tccgcataca
     6781 gctcgataat cttttcaggg ctttgttcat cttcatactc ttccgagcaa aggacgccat
     6841 cggcctcact catgagcaga ttgctccagc catcatgccg ttcaaagtgc aggacctttg
     6901 gaacaggcag ctttccttcc agccatagca tcatgtcctt ttcccgttcc acatcatagg
     6961 tggtcccttt ataccggctg tccgtcattt ttaaatatag gttttcattt tctcccacca
     7021 gcttatatac cttagcagga gacattcctt ccgtatcttt tacgcagcgg tatttttcga
     7081 tcagtttttt caattccggt gatattctca ttttagccat actcttcctt tttcaatatt
     7141 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     7201 aaaataaaca aataggggtt ccgcgcactc ggagccttga tgtgggcgct ttttgcagct
     7261 gttcggctgt gcgctggcca gacagttatg cacaggccag gcgggtttta agagttttaa
     7321 taagttttaa agagttttag gcggaaaaat cgcctttttt ctcttttata tcagtcactt
     7381 acatgtgtga ccggttccca atgtacggct ttgggttccc aatgtacggg ttccggttcc
     7441 caatgtacgg ctttgggttc ccaatgtacg tgctatccac aggaaagaag ctttttcgac
     7501 ctttttcccc tgctagggca atttgcccta gcatctgctc cgtacattag gaaccggcgg
     7561 atgcttcgcc ctcgatcagg ttgcggtagc gcatgactag gatcgggcca gcctgccccg
     7621 cctcctcctt caaatcgtac tccggcaggt catttgaccc gatcagcttg cgcacggtga
     7681 aacagaactt cttgaactct ccggcgctgc cactgcgttc gtagatcgtc ttgaacaacc
     7741 atctggcttc tgccttgcct gcggcgcggc gtgccaggcg gtagagaaaa cggccgatgc
     7801 cgggatcgat caaaaagtaa tcggggtgaa ccgtcagcac gtccgggttc ttgccttctg
     7861 tgatctcgcg gtacatccaa tcagctagct cgatctcgat gtactccggc cgcccggttt
     7921 cgctctttac gatcttgtag cggctaatca aggcttcacc ctcggatacc gtcaccaggc
     7981 ggccgttctt ggccttcttc gtacgctgca tggcaacgtg cgtggtgttt aaccgaatgc
     8041 aggtttctac caggtcgtct ttctgctttc cgccatcggc tcgccggcag aacttgagta
     8101 cgtccgcaac gtgtggacgg aacacgcggc cgggcttgtc tcccttccct tcccggtatc
     8161 ggttcatgga ttcggttaga tgggaaaccg ccatcagtac caggtcgtaa tcccacacac
     8221 tggccatgcc ggccggccct gcggaaacct ctacgtgccc gtctggaagc tcgtagcgga
     8281 tcacctcgcc agctcgtcgg tcacgcttcg acagacggaa aacggccacg tccatgatgc
     8341 tgcgactatc gcgggtgccc acgtcataga gcatcggaac gaaaaaatct ggttgctcgt
     8401 cgcccttggg cggcttccta atcgacgacg caccggctgc cggcggttgc cgggattctt
     8461 tgcggattcg atcagcggcc gcttgccacg attcaccggg gcgtgcttct gcctcgatgc
     8521 gttgccgctg ggcggcctgc gcggccttca acttctccac caggtcatca cccagcgccg
     8581 cgccgatttg taccgggccg gatggtttgc gaccgctcac gccgattcct cgggcttggg
     8641 ggttccagtg ccattgcagg gccggcagac aacccagccg cttacgcctg gccaaccgcc
     8701 cgttcctcca cacatggggc attccacggc gtcggtgcct ggttgttctt gattttccat
     8761 gccgcctcct ttagccgcta aaattcatct actcatttat tcatttgctc atttactctg
     8821 gtagctgcgc gatgtattca gatagcagct cggtaatggt cttgccttgg cgtaccgcgt
     8881 acatcttcag cttggtgtga tcctccgccg gcaactgaaa gttgacccgc ttcatggctg
     8941 gcgtgtctgc caggctggcc aacgttgcag ccttgctgct gcgtgcgctc ggacggccgg
     9001 cacttagcgt gtttgtgctt ttgctcattt tctctttacc tcattaactc aaatgagttt
     9061 tgatttaatt tcagcggcca gcgcctggac ctcgcgggca gcgtcgccct cgggttctga
     9121 ttcaagaacg gttgtgccgg cggcggcagt gcctgggtag ctcacgcgct gcgtgatacg
     9181 ggactcaaga atgggcagct cgtacccggc cagcgcctcg gcaacctcac cgccgatgcg
     9241 cgtgcctttg atcgcccgcg acacgacaaa ggccgcttgt agccttccat ccgtgacctc
     9301 aatg
//
