LOCUS       sequence-342135-        8251 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 8251)
  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 8251)
  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_04085
FEATURES             Location/Qualifiers
     source          1..8251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    771..795
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(862..1569)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1835..2180)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1872..1892)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3213..3465)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4065..4247)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4101..4279)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4442..4466
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4602..5190)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4682..4701)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5361..6155)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6156..6260)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6322..6516
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6582..7655)
                     /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(8084..8251,1..462))
                     /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 ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc
       61 ttgtagcctt ccatccgtga cctcaatgcg ctgcttaacc agctccacca ggtcggcggt
      121 ggcccatatg tcgtaagggc ttggctgcac cggaatcagc acgaagtcgg ctgccttgat
      181 cgcggacaca gccaagtccg ccgcctgggg cgctccgtcg atcactacga agtcgcgccg
      241 gccgatggcc ttcacgtcgc ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg
      301 ttgatcttcc cgcacggccg cccaatcgcg ggcactgccc tggggatcgg aatcgactaa
      361 cagaacatcg gccccggcga gttgcagggc gcgggctaga tgggttgcga tggtcgtctt
      421 gcctgacccg cctttctggt taagtacagc gataaccttc atgcgttccc cttgcgtatt
      481 tgtttattta ctcatcgcat catatacgca gcgaccgcat gacgcaagct gttttactca
      541 aatacacatc acctttttag acggcggcgc tcggtttctt cagcggccaa gctggccggc
      601 caggccgcca gcttggcatc agacaaaccg gccaggattt catgcagccg cacggctgac
      661 tcccgagggg aaccctgtgg ttggcatgca catacaaatg gacgaacgga taaacctttt
      721 cacgcccttt taaatatccg attattctaa taaacgctct tttctcttag gtttacccgc
      781 caatatatcc tgtcaaacac tgatagttta aacggatcac ttcgtgtccc tcggtcacat
      841 gtgcatcctc tagtagcggt cctgctgagc ctcgacatgt tgtcgcaaaa ttcgccctgg
      901 acccgcccaa cgatttgtcg tcactgtcaa ggtttgacct gcacttcatt tggggcccac
      961 atacaccaaa aaaatgctgc ataattctcg gggcagcaag tcggttaccc ggccgccgtg
     1021 ctggaccggg ttgaatggtg cccgtaactt tcggtagagc ggacggccaa tactcaactt
     1081 caaggaatct cacccatgcg cgccggcggg gaaccggagt tcccttcagt gagcgttatt
     1141 agttcgccgc tcggtgtgtc gtagatacta gcccctgggg cacttttgaa atttgaataa
     1201 gatttatgta atcagtcttt taggtttgac cggttctgcc gcttttttta aaattggatt
     1261 tgtaataata aaacgcaatt gtttgttatt gtggcgctct atcatagatg tcgctataaa
     1321 cctattcagc acaatatatt gttttcattt taatattgta catataagta gtagggtaca
     1381 atcagtaaat tgaacggaga atattattca taaaaatacg atagtaacgg gtgatatatt
     1441 cattagaatg aaccgaaacc ggcggtaagg atctgagcta cacatgctca ggttttttac
     1501 aacgtgcaca acagaattga aagcaaatat catgcgatca taggcttctc gcatatctca
     1561 ttaaagcagg acaagctcaa gacttagtca tcttcttctt agccttctta gcagcctcga
     1621 tcaacttaga gatcttgatg tcaggcttct tgatgatctt gttagtagaa gggatgatgt
     1681 tcttccagaa tcctccgtaa ccatcacccc ttccagactg gatcacctcc tgagcctcca
     1741 ccattgtatc gataattgta aatgtaattg taatgttgtt tgttgtttgt tgttgttggt
     1801 aattgttgta aaaatgagct cttatactcg agcgtgtcct ctccaaatga aatgaacttc
     1861 cttatataga ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag
     1921 tggagatgtc acatcaatcc acttgctttg tagacgtggt tggaacctct tctttttcca
     1981 cgatgctcct cgtgggtggg ggtccatctt tgggaccact gtcggcagag agatcttgaa
     2041 tgatagcctt tcctttatcg caatgatggc atttgtagga gccaccttcc ttttctactg
     2101 tcctttcgat gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgaaatta
     2161 tcctttgttg aaaagtctca atagcccttt gatcttctga gactgtatct ttgacatttt
     2221 tggagtagac cagagtgtcg tgctccacca tgttgacgaa gattttcttc ttgtcattga
     2281 gtcgtaaaag actctgtatg aactgttcgc cagtcttgac ggcgagttct gttagatcct
     2341 cgatttgaat cttagactcc atgcatggcc ttagattcag taggaactac ctttttagag
     2401 actccaatct ctattacttg ccttggttta tgaagcaagc cttgaatcgt ccatactgga
     2461 atagtacttc tgatcttgag aaatatgtct ttctctgtgt tcttgatgca attagtcctg
     2521 aatcttttga ctgcatcttt aaccttcttg ggaaggtatt tgatctcctg gagattgtta
     2581 ctcgggtaga tcgtcttgat gagacttgct gcgtaggcct ctctaaccat ctgtgggtca
     2641 gcattctttc tgaaattgaa gaggctaacc ttctcattat cagtggtgaa catagtgtcg
     2701 tcaccttcac cttcgaactt ccttcctaga tcgtaaagat agaggaaatc gtccattgta
     2761 atctccgggg caaaggagat ctcttttggg gctggatcac tgctgggcct tttggttcct
     2821 agcgtgagcc agtgggcttt ttgctttggt gggcttgtta gggccttagc aaagctcttg
     2881 ggcttgagtt gagcttctcc tttggggatg aagttcaacc tgtctgtttg ctgacttgtt
     2941 gtgtacgcgt cagctgctgc tcttgcctct gtaatagtgg caaatttctt gtgtgcaact
     3001 ccgggaacgc cgtttgttgc cgcctttgta caaccccagt catcgtatat accggcatgt
     3061 ggaccgttat acacaacgta gtagttgata tgagggtgtt gaatacccga ttctgctctg
     3121 agaggagcaa ctgtgctgtt aagctcagat ttttgtggga ttggaattcc tccaagcttg
     3181 aattcttgca gcgctccaat tcagcaagcg tcgatctagt aacatagatg acaccgcgcg
     3241 cgataattta tcctagtttg cgcgctatat tttgttttct atcgcgtatt aaatgtataa
     3301 ttgcgggact ctaatcataa aaacccatct cataaataac gtcatgcatt acatgttaat
     3361 tattacatgc ttaacgtaat tcaacagaaa ttatatgata atcatcgcaa gaccggcaac
     3421 aggattcaat cttaagaaac tttattgcca aatgtttgaa cgatctgctt gactctagat
     3481 ccagagtccc gctcaaatct cagtcacagg aagaacaggt ctaggaggaa caggaagaga
     3541 gaaatcaagc tgccagaatc ccacatcatg ccaatttcca tgtttgaaac cagcagctct
     3601 aagcattccc ctaggagcat aaccaagagc ctcatgcatc ctcacagaag gatcattagg
     3661 aagtccaatc acagcaacca cagacttgaa accctgagcc tccaaagact tcaaaagatg
     3721 agtgtaaaga gtagatccaa gaccagttct ttgatgccta ggagacacgt aaacagtaga
     3781 ctcagcagtc caatcgtaag cattcctagc cttccaagga ccagcataag caataccagc
     3841 cacttcacca tccacttcag caacaagcca aggatacctc tctctaagcc tcacaagatc
     3901 atcagtccac tcttgtggtt cttgaggctc agtcctgaaa ttcacagtag aagtctcaat
     3961 gtaatgattc acaatagtgc acacagcagg catatcagcc tcagtagccc ttctaatatc
     4021 agcaggcctt ctttcaggag acatgcgaaa cgatccagat ccggtgcaga ttatttggat
     4081 tgagagtgaa tatgagactc taattggata ccgaggggaa tttatggaac gtcagtggag
     4141 cattttgaca agaaatattt gctagctgat agtgacctta ggcgactttt gaacgcgcaa
     4201 taatggtttc tgacgtatgt gcttagctca ttaaactcca gaaacccgcg gctgagtggc
     4261 tccttcaacg ttgcggttct gtcagttcca aacgtaaaac ggcttgtccc gcgtcatcgg
     4321 cgggggtcat aacgtgactc ccttaattct ccgctccgga tccgaattcg gcacagagtg
     4381 ttcaacccca gtacattaaa aacgtccgca atgtgttatt aagttgtcta agcgtcaatt
     4441 tgtttacacc acaatatatc ctgccaccag ccagccaaca gctccccgac cggcagctcg
     4501 gcacaaaatc accactcgat acaggcagcc catcaaccat gactactgtc cggtccaaca
     4561 gatccgagca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     4621 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4681 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4741 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4801 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4861 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4921 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4981 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5041 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5101 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5161 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     5221 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     5281 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     5341 gagtaaactt ggtctgacag ctaaaacaat tcatccagta aaatataata ttttattttc
     5401 tcccaatcag gcttgatccc cagtaagtca aaaaatagct cgacatactg ttcttccccg
     5461 atatcctccc tgagcgaccg gacgcagaag gcaatgtcat accacttgtc cgccctgccg
     5521 cttctcccaa gatcaataaa gccacttact ttgccatctt tcacaaagat gttgctgtct
     5581 cccaggtcgc cgtgggaaaa gacaagttcc tcttcgggct tttccgtctt taaaaaatca
     5641 tacagctcgc gcggatcttt aaatggagtg tcctcttccc agttttcgca atccacatcg
     5701 gccagatcgt tattcagtaa gtaatccaat tcggctaagc ggctgtctaa gctattcgta
     5761 tagggacaat ccgatatgtc gatggagtga aagagcctga tgcactccgc atacagctcg
     5821 ataatctttt cagggctttg ttcatcttca tactcttccg agcaaaggac gccatcggcc
     5881 tcactcatga gcagattgct ccagccatca tgccgttcaa agtgcaggac ctttggaaca
     5941 ggcagctttc cttccagcca tagcatcatg tccttttccc gttccacatc ataggtggtc
     6001 cctttatacc ggctgtccgt catttttaaa tataggtttt cattttctcc caccagctta
     6061 tataccttag caggagacat tccttccgta tcttttacgc agcggtattt ttcgatcagt
     6121 tttttcaatt ccggtgatat tctcatttta gccatactct tcctttttca atattattga
     6181 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6241 aaacaaatag gggttccgcg cactcggagc cttgatgtgg gcgctttttg cagctgttcg
     6301 gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt tttaataagt
     6361 tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt cacttacatg
     6421 tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg gttcccaatg
     6481 tacggctttg ggttcccaat gtacgtgcta tccacaggaa agaagctttt tcgacctttt
     6541 tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc ggcggatgct
     6601 tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg ccccgcctcc
     6661 tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac ggtgaaacag
     6721 aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa caaccatctg
     6781 gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc gatgccggga
     6841 tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc ttctgtgatc
     6901 tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc ggtttcgctc
     6961 tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac caggcggccg
     7021 ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg aatgcaggtt
     7081 tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt gagtacgtcc
     7141 gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg gtatcggttc
     7201 atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca cacactggcc
     7261 atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta gcggatcacc
     7321 tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat gatgctgcga
     7381 ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg ctcgtcgccc
     7441 ttgggcggct tcctaatcga cgacgcaccg gctgccggcg gttgccggga ttctttgcgg
     7501 attcgatcag cggccgcttg ccacgattca ccggggcgtg cttctgcctc gatgcgttgc
     7561 cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt catcacccag cgccgcgccg
     7621 atttgtaccg ggccggatgg tttgcgaccg ctcacgccga ttcctcgggc ttgggggttc
     7681 cagtgccatt gcagggccgg cagacaaccc agccgcttac gcctggccaa ccgcccgttc
     7741 ctccacacat ggggcattcc acggcgtcgg tgcctggttg ttcttgattt tccatgccgc
     7801 ctcctttagc cgctaaaatt catctactca tttattcatt tgctcattta ctctggtagc
     7861 tgcgcgatgt attcagatag cagctcggta atggtcttgc cttggcgtac cgcgtacatc
     7921 ttcagcttgg tgtgatcctc cgccggcaac tgaaagttga cccgcttcat ggctggcgtg
     7981 tctgccaggc tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg gccggcactt
     8041 agcgtgtttg tgcttttgct cattttctct ttacctcatt aactcaaatg agttttgatt
     8101 taatttcagc ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt tctgattcaa
     8161 gaacggttgt gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg atacgggact
     8221 caagaatggg cagctcgtac ccggccagcg c
//
