LOCUS       sequence-342788-        8251 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 8251)
  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 8251)
  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_18683
FEATURES             Location/Qualifiers
     source          1..8251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    590..614
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(681..1388)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1654..1999)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1691..1711)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3032..3284)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(3884..4066)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(3920..4098)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4261..4285
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4421..5009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4501..4520)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5180..5974)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(5975..6079)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6141..6335
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6401..7474)
                     /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(7903..8251,1..281))
                     /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 gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg
       61 ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt
      121 tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga atcgactaac
      181 agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg
      241 cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc ttgcgtattt
      301 gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg ttttactcaa
      361 atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag ctggccggcc
      421 aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc acggctgact
      481 cccgagggga accctgtggt tggcatgcac atacaaatgg acgaacggat aaaccttttc
      541 acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg tttacccgcc
      601 aatatatcct gtcaaacact gatagtttaa acggatcact tcgtgtccct cggtcacatg
      661 tgcatcctct agtagcggtc ctgctgagcc tcgacatgtt gtcgcaaaat tcgccctgga
      721 cccgcccaac gatttgtcgt cactgtcaag gtttgacctg cacttcattt ggggcccaca
      781 tacaccaaaa aaatgctgca taattctcgg ggcagcaagt cggttacccg gccgccgtgc
      841 tggaccgggt tgaatggtgc ccgtaacttt cggtagagcg gacggccaat actcaacttc
      901 aaggaatctc acccatgcgc gccggcgggg aaccggagtt cccttcagtg agcgttatta
      961 gttcgccgct cggtgtgtcg tagatactag cccctggggc acttttgaaa tttgaataag
     1021 atttatgtaa tcagtctttt aggtttgacc ggttctgccg ctttttttaa aattggattt
     1081 gtaataataa aacgcaattg tttgttattg tggcgctcta tcatagatgt cgctataaac
     1141 ctattcagca caatatattg ttttcatttt aatattgtac atataagtag tagggtacaa
     1201 tcagtaaatt gaacggagaa tattattcat aaaaatacga tagtaacggg tgatatattc
     1261 attagaatga accgaaaccg gcggtaagga tctgagctac acatgctcag gttttttaca
     1321 acgtgcacaa cagaattgaa agcaaatatc atgcgatcat aggcttctcg catatctcat
     1381 taaagcagga caagctcaag acttagtcat cttcttctta gccttcttag cagcctcgat
     1441 caactttccg atcttgatgt caggcttctt gatgatcttg ttagtagaag ggatgatgtt
     1501 cttccagaat cctccgtatc cgtcacccct tccagactgg atcacctcct gagcctccac
     1561 cattgtatcg ataattgtaa atgtaattgt aatgttgttt gttgtttgtt gttgttggta
     1621 attgttgtaa aaatgagctc ttatactcga gcgtgtcctc tccaaatgaa atgaacttcc
     1681 ttatatagag gaagggtctt gcgaaggata gtgggattgt gcgtcatccc ttacgtcagt
     1741 ggagatgtca catcaatcca cttgctttgt agacgtggtt ggaacctctt ctttttccac
     1801 gatgctcctc gtgggtgggg gtccatcttt gggaccactg tcggcagaga gatcttgaat
     1861 gatagccttt cctttatcgc aatgatggca tttgtaggag ccaccttcct tttctactgt
     1921 cctttcgatg aagtgacaga tagctgggca atggaatccg aggaggtttc ccgaaattat
     1981 cctttgttga aaagtctcaa tagccctttg atcttctgag actgtatctt tgacattttt
     2041 ggagtagacc agagtgtcgt gctccaccat gttgacgaag attttcttct tgtcattgag
     2101 tcgtaaaaga ctctgtatga actgttcgcc agtcttgacg gcgagttctg ttagatcctc
     2161 gatttgaatc ttagactcca tgcatggcct tagattcagt aggaactacc tttttagaga
     2221 ctccaatctc tattacttgc cttggtttat gaagcaagcc ttgaatcgtc catactggaa
     2281 tagtacttct gatcttgaga aatatgtctt tctctgtgtt cttgatgcaa ttagtcctga
     2341 atcttttgac tgcatcttta accttcttgg gaaggtattt gatctcctgg agattgttac
     2401 tcgggtagat cgtcttgatg agacttgctg cgtaggcctc tctaaccatc tgtgggtcag
     2461 cattctttct gaaattgaag aggctaacct tctcattatc agtggtgaac atagtgtcgt
     2521 caccttcacc ttcgaacttc cttcctagat cgtaaagata gaggaaatcg tccattgtaa
     2581 tctccggggc aaaggagatc tcttttgggg ctggatcact gctgggcctt ttggttccta
     2641 gcgtgagcca gtgggctttt tgctttggtg ggcttgttag ggccttagca aagctcttgg
     2701 gcttgagttg agcttctcct ttggggatga agttcaacct gtctgtttgc tgacttgttg
     2761 tgtacgcgtc agctgctgct cttgcctctg taatagtggc aaatttcttg tgtgcaactc
     2821 cgggaacgcc gtttgttgcc gcctttgtac aaccccagtc atcgtatata ccggcatgtg
     2881 gaccgttata cacaacgtag tagttgatat gagggtgttg aatacccgat tctgctctga
     2941 gaggagcaac tgtgctgtta agctcagatt tttgtgggat tggaattcct ccaagcttga
     3001 attcttgcag cgctccaatt cagcaagcgt cgatctagta acatagatga caccgcgcgc
     3061 gataatttat cctagtttgc gcgctatatt ttgttttcta tcgcgtatta aatgtataat
     3121 tgcgggactc taatcataaa aacccatctc ataaataacg tcatgcatta catgttaatt
     3181 attacatgct taacgtaatt caacagaaat tatatgataa tcatcgcaag accggcaaca
     3241 ggattcaatc ttaagaaact ttattgccaa atgtttgaac gatctgcttg actctagatc
     3301 cagagtcccg ctcaaatctc agtcacagga agaacaggtc taggaggaac aggaagagag
     3361 aaatcaagct gccagaatcc cacatcatgc caatttccat gtttgaaacc agcagctcta
     3421 agcattcccc taggagcata accaagagcc tcatgcatcc tcacagaagg atcattagga
     3481 agtccaatca cagcaaccac agacttgaaa ccctgagcct ccaaagactt caaaagatga
     3541 gtgtaaagag tagatccaag accagttctt tgatgcctag gagacacgta aacagtagac
     3601 tcagcagtcc aatcgtaagc attcctagcc ttccaaggac cagcataagc aataccagcc
     3661 acttcaccat ccacttcagc aacaagccaa ggatacctct ctctaagcct cacaagatca
     3721 tcagtccact cttgtggttc ttgaggctca gtcctgaaat tcacagtaga agtctcaatg
     3781 taatgattca caatagtgca cacagcaggc atatcagcct cagtagccct tctaatatca
     3841 gcaggccttc tttcaggaga catgcgaaac gatccagatc cggtgcagat tatttggatt
     3901 gagagtgaat atgagactct aattggatac cgaggggaat ttatggaacg tcagtggagc
     3961 attttgacaa gaaatatttg ctagctgata gtgaccttag gcgacttttg aacgcgcaat
     4021 aatggtttct gacgtatgtg cttagctcat taaactccag aaacccgcgg ctgagtggct
     4081 ccttcaacgt tgcggttctg tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc
     4141 gggggtcata acgtgactcc cttaattctc cgctccggat ccgaattcgg cacagagtgt
     4201 tcaaccccag tacattaaaa acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt
     4261 gtttacacca caatatatcc tgccaccagc cagccaacag ctccccgacc ggcagctcgg
     4321 cacaaaatca ccactcgata caggcagccc atcaaccatg actactgtcc ggtccaacag
     4381 atccgagcag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4441 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4501 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4561 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4621 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4681 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4741 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4801 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4861 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4921 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4981 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     5041 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     5101 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     5161 agtaaacttg gtctgacagc taaaacaatt catccagtaa aatataatat tttattttct
     5221 cccaatcagg cttgatcccc agtaagtcaa aaaatagctc gacatactgt tcttccccga
     5281 tatcctccct gagcgaccgg acgcagaagg caatgtcata ccacttgtcc gccctgccgc
     5341 ttctcccaag atcaataaag ccacttactt tgccatcttt cacaaagatg ttgctgtctc
     5401 ccaggtcgcc gtgggaaaag acaagttcct cttcgggctt ttccgtcttt aaaaaatcat
     5461 acagctcgcg cggatcttta aatggagtgt cctcttccca gttttcgcaa tccacatcgg
     5521 ccagatcgtt attcagtaag taatccaatt cggctaagcg gctgtctaag ctattcgtat
     5581 agggacaatc cgatatgtcg atggagtgaa agagcctgat gcactccgca tacagctcga
     5641 taatcttttc agggctttgt tcatcttcat actcttccga gcaaaggacg ccatcggcct
     5701 cactcatgag cagattgctc cagccatcat gccgttcaaa gtgcaggacc tttggaacag
     5761 gcagctttcc ttccagccat agcatcatgt ccttttcccg ttccacatca taggtggtcc
     5821 ctttataccg gctgtccgtc atttttaaat ataggttttc attttctccc accagcttat
     5881 ataccttagc aggagacatt ccttccgtat cttttacgca gcggtatttt tcgatcagtt
     5941 ttttcaattc cggtgatatt ctcattttag ccatactctt cctttttcaa tattattgaa
     6001 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     6061 aacaaatagg ggttccgcgc actcggagcc ttgatgtggg cgctttttgc agctgttcgg
     6121 ctgtgcgctg gccagacagt tatgcacagg ccaggcgggt tttaagagtt ttaataagtt
     6181 ttaaagagtt ttaggcggaa aaatcgcctt ttttctcttt tatatcagtc acttacatgt
     6241 gtgaccggtt cccaatgtac ggctttgggt tcccaatgta cgggttccgg ttcccaatgt
     6301 acggctttgg gttcccaatg tacgtgctat ccacaggaaa gaagcttttt cgaccttttt
     6361 cccctgctag ggcaatttgc cctagcatct gctccgtaca ttaggaaccg gcggatgctt
     6421 cgccctcgat caggttgcgg tagcgcatga ctaggatcgg gccagcctgc cccgcctcct
     6481 ccttcaaatc gtactccggc aggtcatttg acccgatcag cttgcgcacg gtgaaacaga
     6541 acttcttgaa ctctccggcg ctgccactgc gttcgtagat cgtcttgaac aaccatctgg
     6601 cttctgcctt gcctgcggcg cggcgtgcca ggcggtagag aaaacggccg atgccgggat
     6661 cgatcaaaaa gtaatcgggg tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct
     6721 cgcggtacat ccaatcagct agctcgatct cgatgtactc cggccgcccg gtttcgctct
     6781 ttacgatctt gtagcggcta atcaaggctt caccctcgga taccgtcacc aggcggccgt
     6841 tcttggcctt cttcgtacgc tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt
     6901 ctaccaggtc gtctttctgc tttccgccat cggctcgccg gcagaacttg agtacgtccg
     6961 caacgtgtgg acggaacacg cggccgggct tgtctccctt cccttcccgg tatcggttca
     7021 tggattcggt tagatgggaa accgccatca gtaccaggtc gtaatcccac acactggcca
     7081 tgccggccgg ccctgcggaa acctctacgt gcccgtctgg aagctcgtag cggatcacct
     7141 cgccagctcg tcggtcacgc ttcgacagac ggaaaacggc cacgtccatg atgctgcgac
     7201 tatcgcgggt gcccacgtca tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct
     7261 tgggcggctt cctaatcgac gacgcaccgg ctgccggcgg ttgccgggat tctttgcgga
     7321 ttcgatcagc ggccgcttgc cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc
     7381 gctgggcggc ctgcgcggcc ttcaacttct ccaccaggtc atcacccagc gccgcgccga
     7441 tttgtaccgg gccggatggt ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc
     7501 agtgccattg cagggccggc agacaaccca gccgcttacg cctggccaac cgcccgttcc
     7561 tccacacatg gggcattcca cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc
     7621 tcctttagcc gctaaaattc atctactcat ttattcattt gctcatttac tctggtagct
     7681 gcgcgatgta ttcagatagc agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct
     7741 tcagcttggt gtgatcctcc gccggcaact gaaagttgac ccgcttcatg gctggcgtgt
     7801 ctgccaggct ggccaacgtt gcagccttgc tgctgcgtgc gctcggacgg ccggcactta
     7861 gcgtgtttgt gcttttgctc attttctctt tacctcatta actcaaatga gttttgattt
     7921 aatttcagcg gccagcgcct ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag
     7981 aacggttgtg ccggcggcgg cagtgcctgg gtagctcacg cgctgcgtga tacgggactc
     8041 aagaatgggc agctcgtacc cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc
     8101 tttgatcgcc cgcgacacga caaaggccgc ttgtagcctt ccatccgtga cctcaatgcg
     8161 ctgcttaacc agctccacca ggtcggcggt ggcccatatg tcgtaagggc ttggctgcac
     8221 cggaatcagc acgaagtcgg ctgccttgat c
//
