LOCUS       sequence-342113-        8302 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 8302)
  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 8302)
  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_02860
FEATURES             Location/Qualifiers
     source          1..8302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    937..961
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1028..1735)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2052..2397)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2089..2109)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3430..3682)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4282..4464)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4318..4496)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4659..4683
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4819..5407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4899..4918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5578..6372)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6373..6477)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6539..6733
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6799..7872)
                     /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(8301..8302,1..628))
                     /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 aaatgagttt tgatttaatt tcagcggcca gcgcctggac ctcgcgggca gcgtcgccct
       61 cgggttctga ttcaagaacg gttgtgccgg cggcggcagt gcctgggtag ctcacgcgct
      121 gcgtgatacg ggactcaaga atgggcagct cgtacccggc cagcgcctcg gcaacctcac
      181 cgccgatgcg cgtgcctttg atcgcccgcg acacgacaaa ggccgcttgt agccttccat
      241 ccgtgacctc aatgcgctgc ttaaccagct ccaccaggtc ggcggtggcc catatgtcgt
      301 aagggcttgg ctgcaccgga atcagcacga agtcggctgc cttgatcgcg gacacagcca
      361 agtccgccgc ctggggcgct ccgtcgatca ctacgaagtc gcgccggccg atggccttca
      421 cgtcgcggtc aatcgtcggg cggtcgatgc cgacaacggt tagcggttga tcttcccgca
      481 cggccgccca atcgcgggca ctgccctggg gatcggaatc gactaacaga acatcggccc
      541 cggcgagttg cagggcgcgg gctagatggg ttgcgatggt cgtcttgcct gacccgcctt
      601 tctggttaag tacagcgata accttcatgc gttccccttg cgtatttgtt tatttactca
      661 tcgcatcata tacgcagcga ccgcatgacg caagctgttt tactcaaata cacatcacct
      721 ttttagacgg cggcgctcgg tttcttcagc ggccaagctg gccggccagg ccgccagctt
      781 ggcatcagac aaaccggcca ggatttcatg cagccgcacg gctgactccc gaggggaacc
      841 ctgtggttgg catgcacata caaatggacg aacggataaa ccttttcacg cccttttaaa
      901 tatccgatta ttctaataaa cgctcttttc tcttaggttt acccgccaat atatcctgtc
      961 aaacactgat agtttaaacg gatcacttcg tgtccctcgg tcacatgtgc atcctctagt
     1021 agcggtcctg ctgagcctcg acatgttgtc gcaaaattcg ccctggaccc gcccaacgat
     1081 ttgtcgtcac tgtcaaggtt tgacctgcac ttcatttggg gcccacatac accaaaaaaa
     1141 tgctgcataa ttctcggggc agcaagtcgg ttacccggcc gccgtgctgg accgggttga
     1201 atggtgcccg taactttcgg tagagcggac ggccaatact caacttcaag gaatctcacc
     1261 catgcgcgcc ggcggggaac cggagttccc ttcagtgagc gttattagtt cgccgctcgg
     1321 tgtgtcgtag atactagccc ctggggcact tttgaaattt gaataagatt tatgtaatca
     1381 gtcttttagg tttgaccggt tctgccgctt tttttaaaat tggatttgta ataataaaac
     1441 gcaattgttt gttattgtgg cgctctatca tagatgtcgc tataaaccta ttcagcacaa
     1501 tatattgttt tcattttaat attgtacata taagtagtag ggtacaatca gtaaattgaa
     1561 cggagaatat tattcataaa aatacgatag taacgggtga tatattcatt agaatgaacc
     1621 gaaaccggcg gtaaggatct gagctacaca tgctcaggtt ttttacaacg tgcacaacag
     1681 aattgaaagc aaatatcatg cgatcatagg cttctcgcat atctcattaa agcaggacaa
     1741 gctcaaggtc ccctgaaagc ctgagggtcc ttcaagtact tagtgtacaa gttagagatc
     1801 ctagcagcag tgtcgatagg gtatccagac ctgaagatct cgtcgtacac gtcgaaagcc
     1861 tgagacttct tctgcaagaa agcagagaac attgatgaaa gctccttgat agcagccacc
     1921 tcgtcgtcgt ccaacctgaa caactcagac aacttagtca ttgtatcgat aattgtaaat
     1981 gtaattgtaa tgttgtttgt tgtttgttgt tgttggtaat tgttgtaaaa atgagctctt
     2041 atactcgagc gtgtcctctc caaatgaaat gaacttcctt atatagagga agggtcttgc
     2101 gaaggatagt gggattgtgc gtcatccctt acgtcagtgg agatgtcaca tcaatccact
     2161 tgctttgtag acgtggttgg aacctcttct ttttccacga tgctcctcgt gggtgggggt
     2221 ccatctttgg gaccactgtc ggcagagaga tcttgaatga tagcctttcc tttatcgcaa
     2281 tgatggcatt tgtaggagcc accttccttt tctactgtcc tttcgatgaa gtgacagata
     2341 gctgggcaat ggaatccgag gaggtttccc gaaattatcc tttgttgaaa agtctcaata
     2401 gccctttgat cttctgagac tgtatctttg acatttttgg agtagaccag agtgtcgtgc
     2461 tccaccatgt tgacgaagat tttcttcttg tcattgagtc gtaaaagact ctgtatgaac
     2521 tgttcgccag tcttgacggc gagttctgtt agatcctcga tttgaatctt agactccatg
     2581 catggcctta gattcagtag gaactacctt tttagagact ccaatctcta ttacttgcct
     2641 tggtttatga agcaagcctt gaatcgtcca tactggaata gtacttctga tcttgagaaa
     2701 tatgtctttc tctgtgttct tgatgcaatt agtcctgaat cttttgactg catctttaac
     2761 cttcttggga aggtatttga tctcctggag attgttactc gggtagatcg tcttgatgag
     2821 acttgctgcg taggcctctc taaccatctg tgggtcagca ttctttctga aattgaagag
     2881 gctaaccttc tcattatcag tggtgaacat agtgtcgtca ccttcacctt cgaacttcct
     2941 tcctagatcg taaagataga ggaaatcgtc cattgtaatc tccggggcaa aggagatctc
     3001 ttttggggct ggatcactgc tgggcctttt ggttcctagc gtgagccagt gggctttttg
     3061 ctttggtggg cttgttaggg ccttagcaaa gctcttgggc ttgagttgag cttctccttt
     3121 ggggatgaag ttcaacctgt ctgtttgctg acttgttgtg tacgcgtcag ctgctgctct
     3181 tgcctctgta atagtggcaa atttcttgtg tgcaactccg ggaacgccgt ttgttgccgc
     3241 ctttgtacaa ccccagtcat cgtatatacc ggcatgtgga ccgttataca caacgtagta
     3301 gttgatatga gggtgttgaa tacccgattc tgctctgaga ggagcaactg tgctgttaag
     3361 ctcagatttt tgtgggattg gaattcctcc aagcttgaat tcttgcagcg ctccaattca
     3421 gcaagcgtcg atctagtaac atagatgaca ccgcgcgcga taatttatcc tagtttgcgc
     3481 gctatatttt gttttctatc gcgtattaaa tgtataattg cgggactcta atcataaaaa
     3541 cccatctcat aaataacgtc atgcattaca tgttaattat tacatgctta acgtaattca
     3601 acagaaatta tatgataatc atcgcaagac cggcaacagg attcaatctt aagaaacttt
     3661 attgccaaat gtttgaacga tctgcttgac tctagatcca gagtcccgct caaatctcag
     3721 tcacaggaag aacaggtcta ggaggaacag gaagagagaa atcaagctgc cagaatccca
     3781 catcatgcca atttccatgt ttgaaaccag cagctctaag cattccccta ggagcataac
     3841 caagagcctc atgcatcctc acagaaggat cattaggaag tccaatcaca gcaaccacag
     3901 acttgaaacc ctgagcctcc aaagacttca aaagatgagt gtaaagagta gatccaagac
     3961 cagttctttg atgcctagga gacacgtaaa cagtagactc agcagtccaa tcgtaagcat
     4021 tcctagcctt ccaaggacca gcataagcaa taccagccac ttcaccatcc acttcagcaa
     4081 caagccaagg atacctctct ctaagcctca caagatcatc agtccactct tgtggttctt
     4141 gaggctcagt cctgaaattc acagtagaag tctcaatgta atgattcaca atagtgcaca
     4201 cagcaggcat atcagcctca gtagcccttc taatatcagc aggccttctt tcaggagaca
     4261 tgcgaaacga tccagatccg gtgcagatta tttggattga gagtgaatat gagactctaa
     4321 ttggataccg aggggaattt atggaacgtc agtggagcat tttgacaaga aatatttgct
     4381 agctgatagt gaccttaggc gacttttgaa cgcgcaataa tggtttctga cgtatgtgct
     4441 tagctcatta aactccagaa acccgcggct gagtggctcc ttcaacgttg cggttctgtc
     4501 agttccaaac gtaaaacggc ttgtcccgcg tcatcggcgg gggtcataac gtgactccct
     4561 taattctccg ctccggatcc gaattcggca cagagtgttc aaccccagta cattaaaaac
     4621 gtccgcaatg tgttattaag ttgtctaagc gtcaatttgt ttacaccaca atatatcctg
     4681 ccaccagcca gccaacagct ccccgaccgg cagctcggca caaaatcacc actcgataca
     4741 ggcagcccat caaccatgac tactgtccgg tccaacagat ccgagcagga accgtaaaaa
     4801 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4861 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4921 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4981 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5041 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5101 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5161 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5221 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     5281 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     5341 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     5401 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     5461 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     5521 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagcta
     5581 aaacaattca tccagtaaaa tataatattt tattttctcc caatcaggct tgatccccag
     5641 taagtcaaaa aatagctcga catactgttc ttccccgata tcctccctga gcgaccggac
     5701 gcagaaggca atgtcatacc acttgtccgc cctgccgctt ctcccaagat caataaagcc
     5761 acttactttg ccatctttca caaagatgtt gctgtctccc aggtcgccgt gggaaaagac
     5821 aagttcctct tcgggctttt ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa
     5881 tggagtgtcc tcttcccagt tttcgcaatc cacatcggcc agatcgttat tcagtaagta
     5941 atccaattcg gctaagcggc tgtctaagct attcgtatag ggacaatccg atatgtcgat
     6001 ggagtgaaag agcctgatgc actccgcata cagctcgata atcttttcag ggctttgttc
     6061 atcttcatac tcttccgagc aaaggacgcc atcggcctca ctcatgagca gattgctcca
     6121 gccatcatgc cgttcaaagt gcaggacctt tggaacaggc agctttcctt ccagccatag
     6181 catcatgtcc ttttcccgtt ccacatcata ggtggtccct ttataccggc tgtccgtcat
     6241 ttttaaatat aggttttcat tttctcccac cagcttatat accttagcag gagacattcc
     6301 ttccgtatct tttacgcagc ggtatttttc gatcagtttt ttcaattccg gtgatattct
     6361 cattttagcc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6421 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6481 tcggagcctt gatgtgggcg ctttttgcag ctgttcggct gtgcgctggc cagacagtta
     6541 tgcacaggcc aggcgggttt taagagtttt aataagtttt aaagagtttt aggcggaaaa
     6601 atcgcctttt ttctctttta tatcagtcac ttacatgtgt gaccggttcc caatgtacgg
     6661 ctttgggttc ccaatgtacg ggttccggtt cccaatgtac ggctttgggt tcccaatgta
     6721 cgtgctatcc acaggaaaga agctttttcg acctttttcc cctgctaggg caatttgccc
     6781 tagcatctgc tccgtacatt aggaaccggc ggatgcttcg ccctcgatca ggttgcggta
     6841 gcgcatgact aggatcgggc cagcctgccc cgcctcctcc ttcaaatcgt actccggcag
     6901 gtcatttgac ccgatcagct tgcgcacggt gaaacagaac ttcttgaact ctccggcgct
     6961 gccactgcgt tcgtagatcg tcttgaacaa ccatctggct tctgccttgc ctgcggcgcg
     7021 gcgtgccagg cggtagagaa aacggccgat gccgggatcg atcaaaaagt aatcggggtg
     7081 aaccgtcagc acgtccgggt tcttgccttc tgtgatctcg cggtacatcc aatcagctag
     7141 ctcgatctcg atgtactccg gccgcccggt ttcgctcttt acgatcttgt agcggctaat
     7201 caaggcttca ccctcggata ccgtcaccag gcggccgttc ttggccttct tcgtacgctg
     7261 catggcaacg tgcgtggtgt ttaaccgaat gcaggtttct accaggtcgt ctttctgctt
     7321 tccgccatcg gctcgccggc agaacttgag tacgtccgca acgtgtggac ggaacacgcg
     7381 gccgggcttg tctcccttcc cttcccggta tcggttcatg gattcggtta gatgggaaac
     7441 cgccatcagt accaggtcgt aatcccacac actggccatg ccggccggcc ctgcggaaac
     7501 ctctacgtgc ccgtctggaa gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt
     7561 cgacagacgg aaaacggcca cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata
     7621 gagcatcgga acgaaaaaat ctggttgctc gtcgcccttg ggcggcttcc taatcgacga
     7681 cgcaccggct gccggcggtt gccgggattc tttgcggatt cgatcagcgg ccgcttgcca
     7741 cgattcaccg gggcgtgctt ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt
     7801 caacttctcc accaggtcat cacccagcgc cgcgccgatt tgtaccgggc cggatggttt
     7861 gcgaccgctc acgccgattc ctcgggcttg ggggttccag tgccattgca gggccggcag
     7921 acaacccagc cgcttacgcc tggccaaccg cccgttcctc cacacatggg gcattccacg
     7981 gcgtcggtgc ctggttgttc ttgattttcc atgccgcctc ctttagccgc taaaattcat
     8041 ctactcattt attcatttgc tcatttactc tggtagctgc gcgatgtatt cagatagcag
     8101 ctcggtaatg gtcttgcctt ggcgtaccgc gtacatcttc agcttggtgt gatcctccgc
     8161 cggcaactga aagttgaccc gcttcatggc tggcgtgtct gccaggctgg ccaacgttgc
     8221 agccttgctg ctgcgtgcgc tcggacggcc ggcacttagc gtgtttgtgc ttttgctcat
     8281 tttctcttta cctcattaac tc
//
