LOCUS       sequence-342787-        8950 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 8950)
  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 8950)
  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_17309
FEATURES             Location/Qualifiers
     source          1..8950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(239..868)
                     /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    1177..1201
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1268..1975)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2940..3285)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2977..2997)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4318..4570)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5170..5352)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5206..5384)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5547..5571
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5707..6295)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5787..5806)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6466..7260)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7261..7365)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7427..7621
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7687..8760)
                     /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 atttactctg gtagctgcgc gatgtattca gatagcagct cggtaatggt cttgccttgg
       61 cgtaccgcgt acatcttcag cttggtgtga tcctccgccg gcaactgaaa gttgacccgc
      121 ttcatggctg gcgtgtctgc caggctggcc aacgttgcag ccttgctgct gcgtgcgctc
      181 ggacggccgg cacttagcgt gtttgtgctt ttgctcattt tctctttacc tcattaactc
      241 aaatgagttt tgatttaatt tcagcggcca gcgcctggac ctcgcgggca gcgtcgccct
      301 cgggttctga ttcaagaacg gttgtgccgg cggcggcagt gcctgggtag ctcacgcgct
      361 gcgtgatacg ggactcaaga atgggcagct cgtacccggc cagcgcctcg gcaacctcac
      421 cgccgatgcg cgtgcctttg atcgcccgcg acacgacaaa ggccgcttgt agccttccat
      481 ccgtgacctc aatgcgctgc ttaaccagct ccaccaggtc ggcggtggcc catatgtcgt
      541 aagggcttgg ctgcaccgga atcagcacga agtcggctgc cttgatcgcg gacacagcca
      601 agtccgccgc ctggggcgct ccgtcgatca ctacgaagtc gcgccggccg atggccttca
      661 cgtcgcggtc aatcgtcggg cggtcgatgc cgacaacggt tagcggttga tcttcccgca
      721 cggccgccca atcgcgggca ctgccctggg gatcggaatc gactaacaga acatcggccc
      781 cggcgagttg cagggcgcgg gctagatggg ttgcgatggt cgtcttgcct gacccgcctt
      841 tctggttaag tacagcgata accttcatgc gttccccttg cgtatttgtt tatttactca
      901 tcgcatcata tacgcagcga ccgcatgacg caagctgttt tactcaaata cacatcacct
      961 ttttagacgg cggcgctcgg tttcttcagc ggccaagctg gccggccagg ccgccagctt
     1021 ggcatcagac aaaccggcca ggatttcatg cagccgcacg gctgactccc gaggggaacc
     1081 ctgtggttgg catgcacata caaatggacg aacggataaa ccttttcacg cccttttaaa
     1141 tatccgatta ttctaataaa cgctcttttc tcttaggttt acccgccaat atatcctgtc
     1201 aaacactgat agtttaaacg gatcacttcg tgtccctcgg tcacatgtgc atcctctagt
     1261 agcggtcctg ctgagcctcg acatgttgtc gcaaaattcg ccctggaccc gcccaacgat
     1321 ttgtcgtcac tgtcaaggtt tgacctgcac ttcatttggg gcccacatac accaaaaaaa
     1381 tgctgcataa ttctcggggc agcaagtcgg ttacccggcc gccgtgctgg accgggttga
     1441 atggtgcccg taactttcgg tagagcggac ggccaatact caacttcaag gaatctcacc
     1501 catgcgcgcc ggcggggaac cggagttccc ttcagtgagc gttattagtt cgccgctcgg
     1561 tgtgtcgtag atactagccc ctggggcact tttgaaattt gaataagatt tatgtaatca
     1621 gtcttttagg tttgaccggt tctgccgctt tttttaaaat tggatttgta ataataaaac
     1681 gcaattgttt gttattgtgg cgctctatca tagatgtcgc tataaaccta ttcagcacaa
     1741 tatattgttt tcattttaat attgtacata taagtagtag ggtacaatca gtaaattgaa
     1801 cggagaatat tattcataaa aatacgatag taacgggtga tatattcatt agaatgaacc
     1861 gaaaccggcg gtaaggatct gagctacaca tgctcaggtt ttttacaacg tgcacaacag
     1921 aattgaaagc aaatatcatg cgatcatagg cttctcgcat atctcattaa agcaggacaa
     1981 gctcattttc cagaaagcct ttgctcaaga gtctgtgcga tccagtaatt cctgtcgtcc
     2041 tgctttgctg tagcgatcat ctttcccaat ccagcgtcgt cgatacctgc cttcttgaac
     2101 caagacacaa gcaactcgta agggtcattt ctcttcatct tccacacgta gtggaaccag
     2161 atctcaagga tagggttctt caacaatgta gtaccagcat cgttcaattt aagaagttgg
     2221 aacacttcca tctctgtaat atggttcttc ctccatgctg ttgacaactc cttctccaac
     2281 tgtccagaga tcctcttgtt cacacctgac tgagatgaaa cgatcatctt tgccacagtc
     2341 acgtcgtcgt attgctccct caatttcaac aaaagagctt cgatgtaatc tgttgaaggt
     2401 gacttcacgt atgaaagcca tgctggcaat cctggattgt tcaaaacgtt ctgacctcca
     2461 gcgtccaact gcaagatagc gtacacgtcg tcagctgact tacctgactc cttccaagac
     2521 ttctgctgga tctccttcaa ctgatccacc actgtgtccc tagaagtcct cttagcagtt
     2581 gccaacatcc tagccaactc gtcgtcaccg tagtgagcag ccattgtctt gaagatagcc
     2641 ctgtctgcag gcaacctacc cctcttgcag atcttagcca cgtagtgtgc ccagtctgtg
     2701 aactcagaag aagaaaacac gtcagtttta acctgcttaa ccctcaacac ttcaaacctg
     2761 ttgttaacct taagctgctt gtcaagagag aaagtcaact tagcgttgtg tgccaagtcc
     2821 ttaagtgcct tcaccacaga aaaaaccatt gtatcgataa ttgtaaatgt aattgtaatg
     2881 ttgtttgttg tttgttgttg ttggtaattg ttgtaaaaat gagctcttat actcgagcgt
     2941 gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga aggatagtgg
     3001 gattgtgcgt catcccttac gtcagtggag atgtcacatc aatccacttg ctttgtagac
     3061 gtggttggaa cctcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga
     3121 ccactgtcgg cagagagatc ttgaatgata gcctttcctt tatcgcaatg atggcatttg
     3181 taggagccac cttccttttc tactgtcctt tcgatgaagt gacagatagc tgggcaatgg
     3241 aatccgagga ggtttcccga aattatcctt tgttgaaaag tctcaatagc cctttgatct
     3301 tctgagactg tatctttgac atttttggag tagaccagag tgtcgtgctc caccatgttg
     3361 acgaagattt tcttcttgtc attgagtcgt aaaagactct gtatgaactg ttcgccagtc
     3421 ttgacggcga gttctgttag atcctcgatt tgaatcttag actccatgca tggccttaga
     3481 ttcagtagga actacctttt tagagactcc aatctctatt acttgccttg gtttatgaag
     3541 caagccttga atcgtccata ctggaatagt acttctgatc ttgagaaata tgtctttctc
     3601 tgtgttcttg atgcaattag tcctgaatct tttgactgca tctttaacct tcttgggaag
     3661 gtatttgatc tcctggagat tgttactcgg gtagatcgtc ttgatgagac ttgctgcgta
     3721 ggcctctcta accatctgtg ggtcagcatt ctttctgaaa ttgaagaggc taaccttctc
     3781 attatcagtg gtgaacatag tgtcgtcacc ttcaccttcg aacttccttc ctagatcgta
     3841 aagatagagg aaatcgtcca ttgtaatctc cggggcaaag gagatctctt ttggggctgg
     3901 atcactgctg ggccttttgg ttcctagcgt gagccagtgg gctttttgct ttggtgggct
     3961 tgttagggcc ttagcaaagc tcttgggctt gagttgagct tctcctttgg ggatgaagtt
     4021 caacctgtct gtttgctgac ttgttgtgta cgcgtcagct gctgctcttg cctctgtaat
     4081 agtggcaaat ttcttgtgtg caactccggg aacgccgttt gttgccgcct ttgtacaacc
     4141 ccagtcatcg tatataccgg catgtggacc gttatacaca acgtagtagt tgatatgagg
     4201 gtgttgaata cccgattctg ctctgagagg agcaactgtg ctgttaagct cagatttttg
     4261 tgggattgga attcctccaa gcttgaattc ttgcagcgct ccaattcagc aagcgtcgat
     4321 ctagtaacat agatgacacc gcgcgcgata atttatccta gtttgcgcgc tatattttgt
     4381 tttctatcgc gtattaaatg tataattgcg ggactctaat cataaaaacc catctcataa
     4441 ataacgtcat gcattacatg ttaattatta catgcttaac gtaattcaac agaaattata
     4501 tgataatcat cgcaagaccg gcaacaggat tcaatcttaa gaaactttat tgccaaatgt
     4561 ttgaacgatc tgcttgactc tagatccaga gtcccgctca aatctcagtc acaggaagaa
     4621 caggtctagg aggaacagga agagagaaat caagctgcca gaatcccaca tcatgccaat
     4681 ttccatgttt gaaaccagca gctctaagca ttcccctagg agcataacca agagcctcat
     4741 gcatcctcac agaaggatca ttaggaagtc caatcacagc aaccacagac ttgaaaccct
     4801 gagcctccaa agacttcaaa agatgagtgt aaagagtaga tccaagacca gttctttgat
     4861 gcctaggaga cacgtaaaca gtagactcag cagtccaatc gtaagcattc ctagccttcc
     4921 aaggaccagc ataagcaata ccagccactt caccatccac ttcagcaaca agccaaggat
     4981 acctctctct aagcctcaca agatcatcag tccactcttg tggttcttga ggctcagtcc
     5041 tgaaattcac agtagaagtc tcaatgtaat gattcacaat agtgcacaca gcaggcatat
     5101 cagcctcagt agcccttcta atatcagcag gccttctttc aggagacatg cgaaacgatc
     5161 cagatccggt gcagattatt tggattgaga gtgaatatga gactctaatt ggataccgag
     5221 gggaatttat ggaacgtcag tggagcattt tgacaagaaa tatttgctag ctgatagtga
     5281 ccttaggcga cttttgaacg cgcaataatg gtttctgacg tatgtgctta gctcattaaa
     5341 ctccagaaac ccgcggctga gtggctcctt caacgttgcg gttctgtcag ttccaaacgt
     5401 aaaacggctt gtcccgcgtc atcggcgggg gtcataacgt gactccctta attctccgct
     5461 ccggatccga attcggcaca gagtgttcaa ccccagtaca ttaaaaacgt ccgcaatgtg
     5521 ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc accagccagc
     5581 caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg cagcccatca
     5641 accatgacta ctgtccggtc caacagatcc gagcaggaac cgtaaaaagg ccgcgttgct
     5701 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     5761 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5821 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5881 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5941 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6001 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6061 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6121 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     6181 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6241 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     6301 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6361 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     6421 ttttaaatca atctaaagta tatatgagta aacttggtct gacagctaaa acaattcatc
     6481 cagtaaaata taatatttta ttttctccca atcaggcttg atccccagta agtcaaaaaa
     6541 tagctcgaca tactgttctt ccccgatatc ctccctgagc gaccggacgc agaaggcaat
     6601 gtcataccac ttgtccgccc tgccgcttct cccaagatca ataaagccac ttactttgcc
     6661 atctttcaca aagatgttgc tgtctcccag gtcgccgtgg gaaaagacaa gttcctcttc
     6721 gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg gagtgtcctc
     6781 ttcccagttt tcgcaatcca catcggccag atcgttattc agtaagtaat ccaattcggc
     6841 taagcggctg tctaagctat tcgtataggg acaatccgat atgtcgatgg agtgaaagag
     6901 cctgatgcac tccgcataca gctcgataat cttttcaggg ctttgttcat cttcatactc
     6961 ttccgagcaa aggacgccat cggcctcact catgagcaga ttgctccagc catcatgccg
     7021 ttcaaagtgc aggacctttg gaacaggcag ctttccttcc agccatagca tcatgtcctt
     7081 ttcccgttcc acatcatagg tggtcccttt ataccggctg tccgtcattt ttaaatatag
     7141 gttttcattt tctcccacca gcttatatac cttagcagga gacattcctt ccgtatcttt
     7201 tacgcagcgg tatttttcga tcagtttttt caattccggt gatattctca ttttagccat
     7261 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7321 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcactc ggagccttga
     7381 tgtgggcgct ttttgcagct gttcggctgt gcgctggcca gacagttatg cacaggccag
     7441 gcgggtttta agagttttaa taagttttaa agagttttag gcggaaaaat cgcctttttt
     7501 ctcttttata tcagtcactt acatgtgtga ccggttccca atgtacggct ttgggttccc
     7561 aatgtacggg ttccggttcc caatgtacgg ctttgggttc ccaatgtacg tgctatccac
     7621 aggaaagaag ctttttcgac ctttttcccc tgctagggca atttgcccta gcatctgctc
     7681 cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg ttgcggtagc gcatgactag
     7741 gatcgggcca gcctgccccg cctcctcctt caaatcgtac tccggcaggt catttgaccc
     7801 gatcagcttg cgcacggtga aacagaactt cttgaactct ccggcgctgc cactgcgttc
     7861 gtagatcgtc ttgaacaacc atctggcttc tgccttgcct gcggcgcggc gtgccaggcg
     7921 gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa tcggggtgaa ccgtcagcac
     7981 gtccgggttc ttgccttctg tgatctcgcg gtacatccaa tcagctagct cgatctcgat
     8041 gtactccggc cgcccggttt cgctctttac gatcttgtag cggctaatca aggcttcacc
     8101 ctcggatacc gtcaccaggc ggccgttctt ggccttcttc gtacgctgca tggcaacgtg
     8161 cgtggtgttt aaccgaatgc aggtttctac caggtcgtct ttctgctttc cgccatcggc
     8221 tcgccggcag aacttgagta cgtccgcaac gtgtggacgg aacacgcggc cgggcttgtc
     8281 tcccttccct tcccggtatc ggttcatgga ttcggttaga tgggaaaccg ccatcagtac
     8341 caggtcgtaa tcccacacac tggccatgcc ggccggccct gcggaaacct ctacgtgccc
     8401 gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg tcacgcttcg acagacggaa
     8461 aacggccacg tccatgatgc tgcgactatc gcgggtgccc acgtcataga gcatcggaac
     8521 gaaaaaatct ggttgctcgt cgcccttggg cggcttccta atcgacgacg caccggctgc
     8581 cggcggttgc cgggattctt tgcggattcg atcagcggcc gcttgccacg attcaccggg
     8641 gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc gcggccttca acttctccac
     8701 caggtcatca cccagcgccg cgccgatttg taccgggccg gatggtttgc gaccgctcac
     8761 gccgattcct cgggcttggg ggttccagtg ccattgcagg gccggcagac aacccagccg
     8821 cttacgcctg gccaaccgcc cgttcctcca cacatggggc attccacggc gtcggtgcct
     8881 ggttgttctt gattttccat gccgcctcct ttagccgcta aaattcatct actcatttat
     8941 tcatttgctc
//
