LOCUS       sequence-342340-        8377 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 8377)
  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 8377)
  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_13093
FEATURES             Location/Qualifiers
     source          1..8377
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    735..759
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(826..1533)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1925..2270)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1962..1982)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3303..3555)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4155..4337)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4191..4369)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4532..4556
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4692..5280)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4772..4791)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5451..6245)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6246..6350)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6412..6606
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6672..7745)
                     /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(8174..8377,1..426))
                     /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 cgcccgcgac acgacaaagg ccgcttgtag ccttccatcc gtgacctcaa tgcgctgctt
       61 aaccagctcc accaggtcgg cggtggccca tatgtcgtaa gggcttggct gcaccggaat
      121 cagcacgaag tcggctgcct tgatcgcgga cacagccaag tccgccgcct ggggcgctcc
      181 gtcgatcact acgaagtcgc gccggccgat ggccttcacg tcgcggtcaa tcgtcgggcg
      241 gtcgatgccg acaacggtta gcggttgatc ttcccgcacg gccgcccaat cgcgggcact
      301 gccctgggga tcggaatcga ctaacagaac atcggccccg gcgagttgca gggcgcgggc
      361 tagatgggtt gcgatggtcg tcttgcctga cccgcctttc tggttaagta cagcgataac
      421 cttcatgcgt tccccttgcg tatttgttta tttactcatc gcatcatata cgcagcgacc
      481 gcatgacgca agctgtttta ctcaaataca catcaccttt ttagacggcg gcgctcggtt
      541 tcttcagcgg ccaagctggc cggccaggcc gccagcttgg catcagacaa accggccagg
      601 atttcatgca gccgcacggc tgactcccga ggggaaccct gtggttggca tgcacataca
      661 aatggacgaa cggataaacc ttttcacgcc cttttaaata tccgattatt ctaataaacg
      721 ctcttttctc ttaggtttac ccgccaatat atcctgtcaa acactgatag tttaaacgga
      781 tcacttcgtg tccctcggtc acatgtgcat cctctagtag cggtcctgct gagcctcgac
      841 atgttgtcgc aaaattcgcc ctggacccgc ccaacgattt gtcgtcactg tcaaggtttg
      901 acctgcactt catttggggc ccacatacac caaaaaaatg ctgcataatt ctcggggcag
      961 caagtcggtt acccggccgc cgtgctggac cgggttgaat ggtgcccgta actttcggta
     1021 gagcggacgg ccaatactca acttcaagga atctcaccca tgcgcgccgg cggggaaccg
     1081 gagttccctt cagtgagcgt tattagttcg ccgctcggtg tgtcgtagat actagcccct
     1141 ggggcacttt tgaaatttga ataagattta tgtaatcagt cttttaggtt tgaccggttc
     1201 tgccgctttt tttaaaattg gatttgtaat aataaaacgc aattgtttgt tattgtggcg
     1261 ctctatcata gatgtcgcta taaacctatt cagcacaata tattgttttc attttaatat
     1321 tgtacatata agtagtaggg tacaatcagt aaattgaacg gagaatatta ttcataaaaa
     1381 tacgatagta acgggtgata tattcattag aatgaaccga aaccggcggt aaggatctga
     1441 gctacacatg ctcaggtttt ttacaacgtg cacaacagaa ttgaaagcaa atatcatgcg
     1501 atcataggct tctcgcatat ctcattaaag caggacaagc tcagtccaca gtctgaagct
     1561 tagacttcca cctagggtgg ttcttcttcc aaagctccaa gtaagccttc cacaatccgt
     1621 actgagggtt ctctccgata gtcctgtcca acttctgctg agtcaagtac ttgaacacct
     1681 tgtcagggtc gatgttcctc ttcatgatag actgaccgat tccgtcgttg tagatgaatg
     1741 gagactggta cttcacgttt gcagcgtccc acttgtcctt caacacgtcc ttagtcttag
     1801 cgttcaactt ctccacagca gccaaagaag tcattgtatc gataattgta aatgtaattg
     1861 taatgttgtt tgttgtttgt tgttgttggt aattgttgta aaaatgagct cttatactcg
     1921 agcgtgtcct ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat
     1981 agtgggattg tgcgtcatcc cttacgtcag tggagatgtc acatcaatcc acttgctttg
     2041 tagacgtggt tggaacctct tctttttcca cgatgctcct cgtgggtggg ggtccatctt
     2101 tgggaccact gtcggcagag agatcttgaa tgatagcctt tcctttatcg caatgatggc
     2161 atttgtagga gccaccttcc ttttctactg tcctttcgat gaagtgacag atagctgggc
     2221 aatggaatcc gaggaggttt cccgaaatta tcctttgttg aaaagtctca atagcccttt
     2281 gatcttctga gactgtatct ttgacatttt tggagtagac cagagtgtcg tgctccacca
     2341 tgttgacgaa gattttcttc ttgtcattga gtcgtaaaag actctgtatg aactgttcgc
     2401 cagtcttgac ggcgagttct gttagatcct cgatttgaat cttagactcc atgcatggcc
     2461 ttagattcag taggaactac ctttttagag actccaatct ctattacttg ccttggttta
     2521 tgaagcaagc cttgaatcgt ccatactgga atagtacttc tgatcttgag aaatatgtct
     2581 ttctctgtgt tcttgatgca attagtcctg aatcttttga ctgcatcttt aaccttcttg
     2641 ggaaggtatt tgatctcctg gagattgtta ctcgggtaga tcgtcttgat gagacttgct
     2701 gcgtaggcct ctctaaccat ctgtgggtca gcattctttc tgaaattgaa gaggctaacc
     2761 ttctcattat cagtggtgaa catagtgtcg tcaccttcac cttcgaactt ccttcctaga
     2821 tcgtaaagat agaggaaatc gtccattgta atctccgggg caaaggagat ctcttttggg
     2881 gctggatcac tgctgggcct tttggttcct agcgtgagcc agtgggcttt ttgctttggt
     2941 gggcttgtta gggccttagc aaagctcttg ggcttgagtt gagcttctcc tttggggatg
     3001 aagttcaacc tgtctgtttg ctgacttgtt gtgtacgcgt cagctgctgc tcttgcctct
     3061 gtaatagtgg caaatttctt gtgtgcaact ccgggaacgc cgtttgttgc cgcctttgta
     3121 caaccccagt catcgtatat accggcatgt ggaccgttat acacaacgta gtagttgata
     3181 tgagggtgtt gaatacccga ttctgctctg agaggagcaa ctgtgctgtt aagctcagat
     3241 ttttgtggga ttggaattcc tccaagcttg aattcttgca gcgctccaat tcagcaagcg
     3301 tcgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg cgcgctatat
     3361 tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa aaacccatct
     3421 cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat tcaacagaaa
     3481 ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac tttattgcca
     3541 aatgtttgaa cgatctgctt gactctagat ccagagtccc gctcaaatct cagtcacagg
     3601 aagaacaggt ctaggaggaa caggaagaga gaaatcaagc tgccagaatc ccacatcatg
     3661 ccaatttcca tgtttgaaac cagcagctct aagcattccc ctaggagcat aaccaagagc
     3721 ctcatgcatc ctcacagaag gatcattagg aagtccaatc acagcaacca cagacttgaa
     3781 accctgagcc tccaaagact tcaaaagatg agtgtaaaga gtagatccaa gaccagttct
     3841 ttgatgccta ggagacacgt aaacagtaga ctcagcagtc caatcgtaag cattcctagc
     3901 cttccaagga ccagcataag caataccagc cacttcacca tccacttcag caacaagcca
     3961 aggatacctc tctctaagcc tcacaagatc atcagtccac tcttgtggtt cttgaggctc
     4021 agtcctgaaa ttcacagtag aagtctcaat gtaatgattc acaatagtgc acacagcagg
     4081 catatcagcc tcagtagccc ttctaatatc agcaggcctt ctttcaggag acatgcgaaa
     4141 cgatccagat ccggtgcaga ttatttggat tgagagtgaa tatgagactc taattggata
     4201 ccgaggggaa tttatggaac gtcagtggag cattttgaca agaaatattt gctagctgat
     4261 agtgacctta ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt gcttagctca
     4321 ttaaactcca gaaacccgcg gctgagtggc tccttcaacg ttgcggttct gtcagttcca
     4381 aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc ccttaattct
     4441 ccgctccgga tccgaattcg gcacagagtg ttcaacccca gtacattaaa aacgtccgca
     4501 atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc ctgccaccag
     4561 ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat acaggcagcc
     4621 catcaaccat gactactgtc cggtccaaca gatccgagca ggaaccgtaa aaaggccgcg
     4681 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4741 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4801 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4861 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4921 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4981 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5041 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5101 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5161 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5221 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5281 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5341 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5401 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ctaaaacaat
     5461 tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca
     5521 aaaaatagct cgacatactg ttcttccccg atatcctccc tgagcgaccg gacgcagaag
     5581 gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact
     5641 ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc
     5701 tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg
     5761 tcctcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat
     5821 tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga
     5881 aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca
     5941 tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca
     6001 tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg
     6061 tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa
     6121 tataggtttt cattttctcc caccagctta tataccttag caggagacat tccttccgta
     6181 tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta
     6241 gccatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6301 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cactcggagc
     6361 cttgatgtgg gcgctttttg cagctgttcg gctgtgcgct ggccagacag ttatgcacag
     6421 gccaggcggg ttttaagagt tttaataagt tttaaagagt tttaggcgga aaaatcgcct
     6481 tttttctctt ttatatcagt cacttacatg tgtgaccggt tcccaatgta cggctttggg
     6541 ttcccaatgt acgggttccg gttcccaatg tacggctttg ggttcccaat gtacgtgcta
     6601 tccacaggaa agaagctttt tcgacctttt tcccctgcta gggcaatttg ccctagcatc
     6661 tgctccgtac attaggaacc ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg
     6721 actaggatcg ggccagcctg ccccgcctcc tccttcaaat cgtactccgg caggtcattt
     6781 gacccgatca gcttgcgcac ggtgaaacag aacttcttga actctccggc gctgccactg
     6841 cgttcgtaga tcgtcttgaa caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc
     6901 aggcggtaga gaaaacggcc gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc
     6961 agcacgtccg ggttcttgcc ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc
     7021 tcgatgtact ccggccgccc ggtttcgctc tttacgatct tgtagcggct aatcaaggct
     7081 tcaccctcgg ataccgtcac caggcggccg ttcttggcct tcttcgtacg ctgcatggca
     7141 acgtgcgtgg tgtttaaccg aatgcaggtt tctaccaggt cgtctttctg ctttccgcca
     7201 tcggctcgcc ggcagaactt gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc
     7261 ttgtctccct tcccttcccg gtatcggttc atggattcgg ttagatggga aaccgccatc
     7321 agtaccaggt cgtaatccca cacactggcc atgccggccg gccctgcgga aacctctacg
     7381 tgcccgtctg gaagctcgta gcggatcacc tcgccagctc gtcggtcacg cttcgacaga
     7441 cggaaaacgg ccacgtccat gatgctgcga ctatcgcggg tgcccacgtc atagagcatc
     7501 ggaacgaaaa aatctggttg ctcgtcgccc ttgggcggct tcctaatcga cgacgcaccg
     7561 gctgccggcg gttgccggga ttctttgcgg attcgatcag cggccgcttg ccacgattca
     7621 ccggggcgtg cttctgcctc gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc
     7681 tccaccaggt catcacccag cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg
     7741 ctcacgccga ttcctcgggc ttgggggttc cagtgccatt gcagggccgg cagacaaccc
     7801 agccgcttac gcctggccaa ccgcccgttc ctccacacat ggggcattcc acggcgtcgg
     7861 tgcctggttg ttcttgattt tccatgccgc ctcctttagc cgctaaaatt catctactca
     7921 tttattcatt tgctcattta ctctggtagc tgcgcgatgt attcagatag cagctcggta
     7981 atggtcttgc cttggcgtac cgcgtacatc ttcagcttgg tgtgatcctc cgccggcaac
     8041 tgaaagttga cccgcttcat ggctggcgtg tctgccaggc tggccaacgt tgcagccttg
     8101 ctgctgcgtg cgctcggacg gccggcactt agcgtgtttg tgcttttgct cattttctct
     8161 ttacctcatt aactcaaatg agttttgatt taatttcagc ggccagcgcc tggacctcgc
     8221 gggcagcgtc gccctcgggt tctgattcaa gaacggttgt gccggcggcg gcagtgcctg
     8281 ggtagctcac gcgctgcgtg atacgggact caagaatggg cagctcgtac ccggccagcg
     8341 cctcggcaac ctcaccgccg atgcgcgtgc ctttgat
//
