LOCUS       sequence-342203-        8482 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 8482)
  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 8482)
  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_09218
FEATURES             Location/Qualifiers
     source          1..8482
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(282..911)
                     /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    1220..1244
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1311..2018)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2515..2860)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2552..2572)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3893..4145)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4745..4927)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4781..4959)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5122..5146
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5282..5870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5362..5381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6041..6835)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6836..6940)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7002..7196
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7262..8335)
                     /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 cctttagccg ctaaaattca tctactcatt tattcatttg ctcatttact ctggtagctg
       61 cgcgatgtat tcagatagca gctcggtaat ggtcttgcct tggcgtaccg cgtacatctt
      121 cagcttggtg tgatcctccg ccggcaactg aaagttgacc cgcttcatgg ctggcgtgtc
      181 tgccaggctg gccaacgttg cagccttgct gctgcgtgcg ctcggacggc cggcacttag
      241 cgtgtttgtg cttttgctca ttttctcttt acctcattaa ctcaaatgag ttttgattta
      301 atttcagcgg ccagcgcctg gacctcgcgg gcagcgtcgc cctcgggttc tgattcaaga
      361 acggttgtgc cggcggcggc agtgcctggg tagctcacgc gctgcgtgat acgggactca
      421 agaatgggca gctcgtaccc ggccagcgcc tcggcaacct caccgccgat gcgcgtgcct
      481 ttgatcgccc gcgacacgac aaaggccgct tgtagccttc catccgtgac ctcaatgcgc
      541 tgcttaacca gctccaccag gtcggcggtg gcccatatgt cgtaagggct tggctgcacc
      601 ggaatcagca cgaagtcggc tgccttgatc gcggacacag ccaagtccgc cgcctggggc
      661 gctccgtcga tcactacgaa gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc
      721 gggcggtcga tgccgacaac ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg
      781 gcactgccct ggggatcgga atcgactaac agaacatcgg ccccggcgag ttgcagggcg
      841 cgggctagat gggttgcgat ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg
      901 ataaccttca tgcgttcccc ttgcgtattt gtttatttac tcatcgcatc atatacgcag
      961 cgaccgcatg acgcaagctg ttttactcaa atacacatca cctttttaga cggcggcgct
     1021 cggtttcttc agcggccaag ctggccggcc aggccgccag cttggcatca gacaaaccgg
     1081 ccaggatttc atgcagccgc acggctgact cccgagggga accctgtggt tggcatgcac
     1141 atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccga ttattctaat
     1201 aaacgctctt ttctcttagg tttacccgcc aatatatcct gtcaaacact gatagtttaa
     1261 acggatcact tcgtgtccct cggtcacatg tgcatcctct agtagcggtc ctgctgagcc
     1321 tcgacatgtt gtcgcaaaat tcgccctgga cccgcccaac gatttgtcgt cactgtcaag
     1381 gtttgacctg cacttcattt ggggcccaca tacaccaaaa aaatgctgca taattctcgg
     1441 ggcagcaagt cggttacccg gccgccgtgc tggaccgggt tgaatggtgc ccgtaacttt
     1501 cggtagagcg gacggccaat actcaacttc aaggaatctc acccatgcgc gccggcgggg
     1561 aaccggagtt cccttcagtg agcgttatta gttcgccgct cggtgtgtcg tagatactag
     1621 cccctggggc acttttgaaa tttgaataag atttatgtaa tcagtctttt aggtttgacc
     1681 ggttctgccg ctttttttaa aattggattt gtaataataa aacgcaattg tttgttattg
     1741 tggcgctcta tcatagatgt cgctataaac ctattcagca caatatattg ttttcatttt
     1801 aatattgtac atataagtag tagggtacaa tcagtaaatt gaacggagaa tattattcat
     1861 aaaaatacga tagtaacggg tgatatattc attagaatga accgaaaccg gcggtaagga
     1921 tctgagctac acatgctcag gttttttaca acgtgcacaa cagaattgaa agcaaatatc
     1981 atgcgatcat aggcttctcg catatctcat taaagcagga caagctcaag cagatccagt
     2041 agtagttgta gtagttgcag cagttgaaga tttcctgtcg aaaagagcct tgtaagcacc
     2101 gtagatagcg aatccagcca cagtagctcc caaagtcacc tttgcgaact tcctcaacct
     2161 tggccaagac ttagggttct tctgcactgt tccagccatc atagctgaga ttttagcgat
     2221 tccctcgtca gacatcttag cagactcctt agcagcagcg aaagtagtga tagccttcct
     2281 cactgcagcc tcgtcagcgt tagagttctt gatagtgtta gcagccttag tcaaagcagt
     2341 gttagtctta agcttctcca agaatcctcc agccaacttt gccacgttgt ccttgttcac
     2401 aggcacagga gcagcgtcag ccattgtatc gataattgta aatgtaattg taatgttgtt
     2461 tgttgtttgt tgttgttggt aattgttgta aaaatgagct cttatactcg agcgtgtcct
     2521 ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat agtgggattg
     2581 tgcgtcatcc cttacgtcag tggagatgtc acatcaatcc acttgctttg tagacgtggt
     2641 tggaacctct tctttttcca cgatgctcct cgtgggtggg ggtccatctt tgggaccact
     2701 gtcggcagag agatcttgaa tgatagcctt tcctttatcg caatgatggc atttgtagga
     2761 gccaccttcc ttttctactg tcctttcgat gaagtgacag atagctgggc aatggaatcc
     2821 gaggaggttt cccgaaatta tcctttgttg aaaagtctca atagcccttt gatcttctga
     2881 gactgtatct ttgacatttt tggagtagac cagagtgtcg tgctccacca tgttgacgaa
     2941 gattttcttc ttgtcattga gtcgtaaaag actctgtatg aactgttcgc cagtcttgac
     3001 ggcgagttct gttagatcct cgatttgaat cttagactcc atgcatggcc ttagattcag
     3061 taggaactac ctttttagag actccaatct ctattacttg ccttggttta tgaagcaagc
     3121 cttgaatcgt ccatactgga atagtacttc tgatcttgag aaatatgtct ttctctgtgt
     3181 tcttgatgca attagtcctg aatcttttga ctgcatcttt aaccttcttg ggaaggtatt
     3241 tgatctcctg gagattgtta ctcgggtaga tcgtcttgat gagacttgct gcgtaggcct
     3301 ctctaaccat ctgtgggtca gcattctttc tgaaattgaa gaggctaacc ttctcattat
     3361 cagtggtgaa catagtgtcg tcaccttcac cttcgaactt ccttcctaga tcgtaaagat
     3421 agaggaaatc gtccattgta atctccgggg caaaggagat ctcttttggg gctggatcac
     3481 tgctgggcct tttggttcct agcgtgagcc agtgggcttt ttgctttggt gggcttgtta
     3541 gggccttagc aaagctcttg ggcttgagtt gagcttctcc tttggggatg aagttcaacc
     3601 tgtctgtttg ctgacttgtt gtgtacgcgt cagctgctgc tcttgcctct gtaatagtgg
     3661 caaatttctt gtgtgcaact ccgggaacgc cgtttgttgc cgcctttgta caaccccagt
     3721 catcgtatat accggcatgt ggaccgttat acacaacgta gtagttgata tgagggtgtt
     3781 gaatacccga ttctgctctg agaggagcaa ctgtgctgtt aagctcagat ttttgtggga
     3841 ttggaattcc tccaagcttg aattcttgca gcgctccaat tcagcaagcg tcgatctagt
     3901 aacatagatg acaccgcgcg cgataattta tcctagtttg cgcgctatat tttgttttct
     3961 atcgcgtatt aaatgtataa ttgcgggact ctaatcataa aaacccatct cataaataac
     4021 gtcatgcatt acatgttaat tattacatgc ttaacgtaat tcaacagaaa ttatatgata
     4081 atcatcgcaa gaccggcaac aggattcaat cttaagaaac tttattgcca aatgtttgaa
     4141 cgatctgctt gactctagat ccagagtccc gctcaaatct cagtcacagg aagaacaggt
     4201 ctaggaggaa caggaagaga gaaatcaagc tgccagaatc ccacatcatg ccaatttcca
     4261 tgtttgaaac cagcagctct aagcattccc ctaggagcat aaccaagagc ctcatgcatc
     4321 ctcacagaag gatcattagg aagtccaatc acagcaacca cagacttgaa accctgagcc
     4381 tccaaagact tcaaaagatg agtgtaaaga gtagatccaa gaccagttct ttgatgccta
     4441 ggagacacgt aaacagtaga ctcagcagtc caatcgtaag cattcctagc cttccaagga
     4501 ccagcataag caataccagc cacttcacca tccacttcag caacaagcca aggatacctc
     4561 tctctaagcc tcacaagatc atcagtccac tcttgtggtt cttgaggctc agtcctgaaa
     4621 ttcacagtag aagtctcaat gtaatgattc acaatagtgc acacagcagg catatcagcc
     4681 tcagtagccc ttctaatatc agcaggcctt ctttcaggag acatgcgaaa cgatccagat
     4741 ccggtgcaga ttatttggat tgagagtgaa tatgagactc taattggata ccgaggggaa
     4801 tttatggaac gtcagtggag cattttgaca agaaatattt gctagctgat agtgacctta
     4861 ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt gcttagctca ttaaactcca
     4921 gaaacccgcg gctgagtggc tccttcaacg ttgcggttct gtcagttcca aacgtaaaac
     4981 ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc ccttaattct ccgctccgga
     5041 tccgaattcg gcacagagtg ttcaacccca gtacattaaa aacgtccgca atgtgttatt
     5101 aagttgtcta agcgtcaatt tgtttacacc acaatatatc ctgccaccag ccagccaaca
     5161 gctccccgac cggcagctcg gcacaaaatc accactcgat acaggcagcc catcaaccat
     5221 gactactgtc cggtccaaca gatccgagca ggaaccgtaa aaaggccgcg ttgctggcgt
     5281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5461 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5701 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5821 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5881 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5941 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6001 aatcaatcta aagtatatat gagtaaactt ggtctgacag ctaaaacaat tcatccagta
     6061 aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca aaaaatagct
     6121 cgacatactg ttcttccccg atatcctccc tgagcgaccg gacgcagaag gcaatgtcat
     6181 accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact ttgccatctt
     6241 tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc tcttcgggct
     6301 tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg tcctcttccc
     6361 agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat tcggctaagc
     6421 ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga aagagcctga
     6481 tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca tactcttccg
     6541 agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca tgccgttcaa
     6601 agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg tccttttccc
     6661 gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa tataggtttt
     6721 cattttctcc caccagctta tataccttag caggagacat tccttccgta tcttttacgc
     6781 agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta gccatactct
     6841 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6901 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cactcggagc cttgatgtgg
     6961 gcgctttttg cagctgttcg gctgtgcgct ggccagacag ttatgcacag gccaggcggg
     7021 ttttaagagt tttaataagt tttaaagagt tttaggcgga aaaatcgcct tttttctctt
     7081 ttatatcagt cacttacatg tgtgaccggt tcccaatgta cggctttggg ttcccaatgt
     7141 acgggttccg gttcccaatg tacggctttg ggttcccaat gtacgtgcta tccacaggaa
     7201 agaagctttt tcgacctttt tcccctgcta gggcaatttg ccctagcatc tgctccgtac
     7261 attaggaacc ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg actaggatcg
     7321 ggccagcctg ccccgcctcc tccttcaaat cgtactccgg caggtcattt gacccgatca
     7381 gcttgcgcac ggtgaaacag aacttcttga actctccggc gctgccactg cgttcgtaga
     7441 tcgtcttgaa caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga
     7501 gaaaacggcc gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg
     7561 ggttcttgcc ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc tcgatgtact
     7621 ccggccgccc ggtttcgctc tttacgatct tgtagcggct aatcaaggct tcaccctcgg
     7681 ataccgtcac caggcggccg ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg
     7741 tgtttaaccg aatgcaggtt tctaccaggt cgtctttctg ctttccgcca tcggctcgcc
     7801 ggcagaactt gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc ttgtctccct
     7861 tcccttcccg gtatcggttc atggattcgg ttagatggga aaccgccatc agtaccaggt
     7921 cgtaatccca cacactggcc atgccggccg gccctgcgga aacctctacg tgcccgtctg
     7981 gaagctcgta gcggatcacc tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg
     8041 ccacgtccat gatgctgcga ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa
     8101 aatctggttg ctcgtcgccc ttgggcggct tcctaatcga cgacgcaccg gctgccggcg
     8161 gttgccggga ttctttgcgg attcgatcag cggccgcttg ccacgattca ccggggcgtg
     8221 cttctgcctc gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc tccaccaggt
     8281 catcacccag cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg ctcacgccga
     8341 ttcctcgggc ttgggggttc cagtgccatt gcagggccgg cagacaaccc agccgcttac
     8401 gcctggccaa ccgcccgttc ctccacacat ggggcattcc acggcgtcgg tgcctggttg
     8461 ttcttgattt tccatgccgc ct
//
