LOCUS       sequence-342343-        9406 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 9406)
  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 9406)
  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_15039a
FEATURES             Location/Qualifiers
     source          1..9406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(195..824)
                     /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    1133..1157
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1224..1931)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(3352..3697)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(3389..3409)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(4730..4982)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(5582..5764)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(5618..5796)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5959..5983
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6119..6707)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6199..6218)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6878..7672)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(7673..7777)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7839..8033
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(8099..9172)
                     /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 aatggtcttg ccttggcgta ccgcgtacat cttcagcttg gtgtgatcct ccgccggcaa
       61 ctgaaagttg acccgcttca tggctggcgt gtctgccagg ctggccaacg ttgcagcctt
      121 gctgctgcgt gcgctcggac ggccggcact tagcgtgttt gtgcttttgc tcattttctc
      181 tttacctcat taactcaaat gagttttgat ttaatttcag cggccagcgc ctggacctcg
      241 cgggcagcgt cgccctcggg ttctgattca agaacggttg tgccggcggc ggcagtgcct
      301 gggtagctca cgcgctgcgt gatacgggac tcaagaatgg gcagctcgta cccggccagc
      361 gcctcggcaa cctcaccgcc gatgcgcgtg cctttgatcg cccgcgacac gacaaaggcc
      421 gcttgtagcc ttccatccgt gacctcaatg cgctgcttaa ccagctccac caggtcggcg
      481 gtggcccata tgtcgtaagg gcttggctgc accggaatca gcacgaagtc ggctgccttg
      541 atcgcggaca cagccaagtc cgccgcctgg ggcgctccgt cgatcactac gaagtcgcgc
      601 cggccgatgg ccttcacgtc gcggtcaatc gtcgggcggt cgatgccgac aacggttagc
      661 ggttgatctt cccgcacggc cgcccaatcg cgggcactgc cctggggatc ggaatcgact
      721 aacagaacat cggccccggc gagttgcagg gcgcgggcta gatgggttgc gatggtcgtc
      781 ttgcctgacc cgcctttctg gttaagtaca gcgataacct tcatgcgttc cccttgcgta
      841 tttgtttatt tactcatcgc atcatatacg cagcgaccgc atgacgcaag ctgttttact
      901 caaatacaca tcaccttttt agacggcggc gctcggtttc ttcagcggcc aagctggccg
      961 gccaggccgc cagcttggca tcagacaaac cggccaggat ttcatgcagc cgcacggctg
     1021 actcccgagg ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt
     1081 ttcacgccct tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc
     1141 gccaatatat cctgtcaaac actgatagtt taaacggatc acttcgtgtc cctcggtcac
     1201 atgtgcatcc tctagtagcg gtcctgctga gcctcgacat gttgtcgcaa aattcgccct
     1261 ggacccgccc aacgatttgt cgtcactgtc aaggtttgac ctgcacttca tttggggccc
     1321 acatacacca aaaaaatgct gcataattct cggggcagca agtcggttac ccggccgccg
     1381 tgctggaccg ggttgaatgg tgcccgtaac tttcggtaga gcggacggcc aatactcaac
     1441 ttcaaggaat ctcacccatg cgcgccggcg gggaaccgga gttcccttca gtgagcgtta
     1501 ttagttcgcc gctcggtgtg tcgtagatac tagcccctgg ggcacttttg aaatttgaat
     1561 aagatttatg taatcagtct tttaggtttg accggttctg ccgctttttt taaaattgga
     1621 tttgtaataa taaaacgcaa ttgtttgtta ttgtggcgct ctatcataga tgtcgctata
     1681 aacctattca gcacaatata ttgttttcat tttaatattg tacatataag tagtagggta
     1741 caatcagtaa attgaacgga gaatattatt cataaaaata cgatagtaac gggtgatata
     1801 ttcattagaa tgaaccgaaa ccggcggtaa ggatctgagc tacacatgct caggtttttt
     1861 acaacgtgca caacagaatt gaaagcaaat atcatgcgat cataggcttc tcgcatatct
     1921 cattaaagca ggacaagctc atcctgactt cttttccttc aacaatttag ccaatgcatt
     1981 tctgtatttg ttgtaaagaa gcttcctcac gtcgtcctta ccagcaccag acactgaaag
     2041 aaggtagaac accctcactg agtctgtctc agttgccacc cacctgttga tctgtgcgta
     2101 caaaagctgc tcagcgtggt acctagtctt tggaacgtgc catgctctca cgaaagcagt
     2161 agtaagagat ctctcgtcct cataaatttt tgtaagagaa gaaagcaagt cacctccgtc
     2221 ctttggcttc ttacctctgt agtaatcatt aaaccttgcc caagacacga acaaaggatt
     2281 ctcaagcaaa ggagtgttca catcatccaa cttaagcaac tcgaaaacct tctttggagt
     2341 caattttgac tcgtaccagt tcctgatctg ctggatgaac aaatgctgag cgataagaga
     2401 tgacctagga tttttgtctg cagcaacgat catcttagcc aacacgtcgt cagcatagaa
     2461 cttcttcaac actgaaatag gtgtcatttt tgttttgtga tggatcaatg aaaagtcgtc
     2521 agtataccta acccatgcag tgaacaatgg agactccaac aatggcacct tagtctcctg
     2581 caatttcaac aaattgaaaa cgtcgatagg ttgcttctta gttttcaacc aaagagcagt
     2641 ctgctcagct tggatcctaa tagcgatttt ctctgttgaa ggcacgttct tagctgcgat
     2701 aagcatgtgc acgaaagcct cctcgtcttt gtacaaaggt ttcaaaaaag aaaaaagagt
     2761 agtctctgat gtagggttaa ccttcttata ctcgtcaacg aacttagccc atggcacgat
     2821 ctgaggctgt gtgaaaagag actcagctgt gtcctccaac ttcaacaact tgaaaactga
     2881 gtctggtgtc tccttagcca acaaccacct gtagatctgc tcagtctgca ccctttttgc
     2941 caaaccagct gtagccctca cctgcttagc agcctcgatg atcttagcaa gaccgtcatc
     3001 cttgtaatga gaagtcaatg tagcgataac tgttgtcctc ttagtaggat ttgctttatt
     3061 gtacaatttc atataaccaa tccagtcatt caacctagga tttgcaagca agtcgtctgc
     3121 agccttgtcc aacatcaaaa gcttgaacac gtcatctgca gactctccct tcttaagtgc
     3181 gtcgtccaac tgctgtgaca ctgatgattt aagagagttt gtcaaagtct cgatagctga
     3241 tgttccagga gatggaccca ttgtatcgat aattgtaaat gtaattgtaa tgttgtttgt
     3301 tgtttgttgt tgttggtaat tgttgtaaaa atgagctctt atactcgagc gtgtcctctc
     3361 caaatgaaat gaacttcctt atatagagga agggtcttgc gaaggatagt gggattgtgc
     3421 gtcatccctt acgtcagtgg agatgtcaca tcaatccact tgctttgtag acgtggttgg
     3481 aacctcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc
     3541 ggcagagaga tcttgaatga tagcctttcc tttatcgcaa tgatggcatt tgtaggagcc
     3601 accttccttt tctactgtcc tttcgatgaa gtgacagata gctgggcaat ggaatccgag
     3661 gaggtttccc gaaattatcc tttgttgaaa agtctcaata gccctttgat cttctgagac
     3721 tgtatctttg acatttttgg agtagaccag agtgtcgtgc tccaccatgt tgacgaagat
     3781 tttcttcttg tcattgagtc gtaaaagact ctgtatgaac tgttcgccag tcttgacggc
     3841 gagttctgtt agatcctcga tttgaatctt agactccatg catggcctta gattcagtag
     3901 gaactacctt tttagagact ccaatctcta ttacttgcct tggtttatga agcaagcctt
     3961 gaatcgtcca tactggaata gtacttctga tcttgagaaa tatgtctttc tctgtgttct
     4021 tgatgcaatt agtcctgaat cttttgactg catctttaac cttcttggga aggtatttga
     4081 tctcctggag attgttactc gggtagatcg tcttgatgag acttgctgcg taggcctctc
     4141 taaccatctg tgggtcagca ttctttctga aattgaagag gctaaccttc tcattatcag
     4201 tggtgaacat agtgtcgtca ccttcacctt cgaacttcct tcctagatcg taaagataga
     4261 ggaaatcgtc cattgtaatc tccggggcaa aggagatctc ttttggggct ggatcactgc
     4321 tgggcctttt ggttcctagc gtgagccagt gggctttttg ctttggtggg cttgttaggg
     4381 ccttagcaaa gctcttgggc ttgagttgag cttctccttt ggggatgaag ttcaacctgt
     4441 ctgtttgctg acttgttgtg tacgcgtcag ctgctgctct tgcctctgta atagtggcaa
     4501 atttcttgtg tgcaactccg ggaacgccgt ttgttgccgc ctttgtacaa ccccagtcat
     4561 cgtatatacc ggcatgtgga ccgttataca caacgtagta gttgatatga gggtgttgaa
     4621 tacccgattc tgctctgaga ggagcaactg tgctgttaag ctcagatttt tgtgggattg
     4681 gaattcctcc aagcttgaat tcttgcagcg ctccaattca gcaagcgtcg atctagtaac
     4741 atagatgaca ccgcgcgcga taatttatcc tagtttgcgc gctatatttt gttttctatc
     4801 gcgtattaaa tgtataattg cgggactcta atcataaaaa cccatctcat aaataacgtc
     4861 atgcattaca tgttaattat tacatgctta acgtaattca acagaaatta tatgataatc
     4921 atcgcaagac cggcaacagg attcaatctt aagaaacttt attgccaaat gtttgaacga
     4981 tctgcttgac tctagatcca gagtcccgct caaatctcag tcacaggaag aacaggtcta
     5041 ggaggaacag gaagagagaa atcaagctgc cagaatccca catcatgcca atttccatgt
     5101 ttgaaaccag cagctctaag cattccccta ggagcataac caagagcctc atgcatcctc
     5161 acagaaggat cattaggaag tccaatcaca gcaaccacag acttgaaacc ctgagcctcc
     5221 aaagacttca aaagatgagt gtaaagagta gatccaagac cagttctttg atgcctagga
     5281 gacacgtaaa cagtagactc agcagtccaa tcgtaagcat tcctagcctt ccaaggacca
     5341 gcataagcaa taccagccac ttcaccatcc acttcagcaa caagccaagg atacctctct
     5401 ctaagcctca caagatcatc agtccactct tgtggttctt gaggctcagt cctgaaattc
     5461 acagtagaag tctcaatgta atgattcaca atagtgcaca cagcaggcat atcagcctca
     5521 gtagcccttc taatatcagc aggccttctt tcaggagaca tgcgaaacga tccagatccg
     5581 gtgcagatta tttggattga gagtgaatat gagactctaa ttggataccg aggggaattt
     5641 atggaacgtc agtggagcat tttgacaaga aatatttgct agctgatagt gaccttaggc
     5701 gacttttgaa cgcgcaataa tggtttctga cgtatgtgct tagctcatta aactccagaa
     5761 acccgcggct gagtggctcc ttcaacgttg cggttctgtc agttccaaac gtaaaacggc
     5821 ttgtcccgcg tcatcggcgg gggtcataac gtgactccct taattctccg ctccggatcc
     5881 gaattcggca cagagtgttc aaccccagta cattaaaaac gtccgcaatg tgttattaag
     5941 ttgtctaagc gtcaatttgt ttacaccaca atatatcctg ccaccagcca gccaacagct
     6001 ccccgaccgg cagctcggca caaaatcacc actcgataca ggcagcccat caaccatgac
     6061 tactgtccgg tccaacagat ccgagcagga accgtaaaaa ggccgcgttg ctggcgtttt
     6121 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     6181 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     6241 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6301 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6361 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6421 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6481 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6541 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6601 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6661 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6721 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6781 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6841 caatctaaag tatatatgag taaacttggt ctgacagcta aaacaattca tccagtaaaa
     6901 tataatattt tattttctcc caatcaggct tgatccccag taagtcaaaa aatagctcga
     6961 catactgttc ttccccgata tcctccctga gcgaccggac gcagaaggca atgtcatacc
     7021 acttgtccgc cctgccgctt ctcccaagat caataaagcc acttactttg ccatctttca
     7081 caaagatgtt gctgtctccc aggtcgccgt gggaaaagac aagttcctct tcgggctttt
     7141 ccgtctttaa aaaatcatac agctcgcgcg gatctttaaa tggagtgtcc tcttcccagt
     7201 tttcgcaatc cacatcggcc agatcgttat tcagtaagta atccaattcg gctaagcggc
     7261 tgtctaagct attcgtatag ggacaatccg atatgtcgat ggagtgaaag agcctgatgc
     7321 actccgcata cagctcgata atcttttcag ggctttgttc atcttcatac tcttccgagc
     7381 aaaggacgcc atcggcctca ctcatgagca gattgctcca gccatcatgc cgttcaaagt
     7441 gcaggacctt tggaacaggc agctttcctt ccagccatag catcatgtcc ttttcccgtt
     7501 ccacatcata ggtggtccct ttataccggc tgtccgtcat ttttaaatat aggttttcat
     7561 tttctcccac cagcttatat accttagcag gagacattcc ttccgtatct tttacgcagc
     7621 ggtatttttc gatcagtttt ttcaattccg gtgatattct cattttagcc atactcttcc
     7681 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     7741 aatgtattta gaaaaataaa caaatagggg ttccgcgcac tcggagcctt gatgtgggcg
     7801 ctttttgcag ctgttcggct gtgcgctggc cagacagtta tgcacaggcc aggcgggttt
     7861 taagagtttt aataagtttt aaagagtttt aggcggaaaa atcgcctttt ttctctttta
     7921 tatcagtcac ttacatgtgt gaccggttcc caatgtacgg ctttgggttc ccaatgtacg
     7981 ggttccggtt cccaatgtac ggctttgggt tcccaatgta cgtgctatcc acaggaaaga
     8041 agctttttcg acctttttcc cctgctaggg caatttgccc tagcatctgc tccgtacatt
     8101 aggaaccggc ggatgcttcg ccctcgatca ggttgcggta gcgcatgact aggatcgggc
     8161 cagcctgccc cgcctcctcc ttcaaatcgt actccggcag gtcatttgac ccgatcagct
     8221 tgcgcacggt gaaacagaac ttcttgaact ctccggcgct gccactgcgt tcgtagatcg
     8281 tcttgaacaa ccatctggct tctgccttgc ctgcggcgcg gcgtgccagg cggtagagaa
     8341 aacggccgat gccgggatcg atcaaaaagt aatcggggtg aaccgtcagc acgtccgggt
     8401 tcttgccttc tgtgatctcg cggtacatcc aatcagctag ctcgatctcg atgtactccg
     8461 gccgcccggt ttcgctcttt acgatcttgt agcggctaat caaggcttca ccctcggata
     8521 ccgtcaccag gcggccgttc ttggccttct tcgtacgctg catggcaacg tgcgtggtgt
     8581 ttaaccgaat gcaggtttct accaggtcgt ctttctgctt tccgccatcg gctcgccggc
     8641 agaacttgag tacgtccgca acgtgtggac ggaacacgcg gccgggcttg tctcccttcc
     8701 cttcccggta tcggttcatg gattcggtta gatgggaaac cgccatcagt accaggtcgt
     8761 aatcccacac actggccatg ccggccggcc ctgcggaaac ctctacgtgc ccgtctggaa
     8821 gctcgtagcg gatcacctcg ccagctcgtc ggtcacgctt cgacagacgg aaaacggcca
     8881 cgtccatgat gctgcgacta tcgcgggtgc ccacgtcata gagcatcgga acgaaaaaat
     8941 ctggttgctc gtcgcccttg ggcggcttcc taatcgacga cgcaccggct gccggcggtt
     9001 gccgggattc tttgcggatt cgatcagcgg ccgcttgcca cgattcaccg gggcgtgctt
     9061 ctgcctcgat gcgttgccgc tgggcggcct gcgcggcctt caacttctcc accaggtcat
     9121 cacccagcgc cgcgccgatt tgtaccgggc cggatggttt gcgaccgctc acgccgattc
     9181 ctcgggcttg ggggttccag tgccattgca gggccggcag acaacccagc cgcttacgcc
     9241 tggccaaccg cccgttcctc cacacatggg gcattccacg gcgtcggtgc ctggttgttc
     9301 ttgattttcc atgccgcctc ctttagccgc taaaattcat ctactcattt attcatttgc
     9361 tcatttactc tggtagctgc gcgatgtatt cagatagcag ctcggt
//
