LOCUS       sequence-342756-        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_17316
FEATURES             Location/Qualifiers
     source          1..8950
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    751..775
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(842..1549)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2514..2859)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2551..2571)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3892..4144)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4744..4926)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4780..4958)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    5121..5145
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5281..5869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5361..5380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6040..6834)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6835..6939)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7001..7195
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7261..8334)
                     /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(8763..8950,1..442))
                     /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 tgcgcgtgcc tttgatcgcc cgcgacacga caaaggccgc ttgtagcctt ccatccgtga
       61 cctcaatgcg ctgcttaacc agctccacca ggtcggcggt ggcccatatg tcgtaagggc
      121 ttggctgcac cggaatcagc acgaagtcgg ctgccttgat cgcggacaca gccaagtccg
      181 ccgcctgggg cgctccgtcg atcactacga agtcgcgccg gccgatggcc ttcacgtcgc
      241 ggtcaatcgt cgggcggtcg atgccgacaa cggttagcgg ttgatcttcc cgcacggccg
      301 cccaatcgcg ggcactgccc tggggatcgg aatcgactaa cagaacatcg gccccggcga
      361 gttgcagggc gcgggctaga tgggttgcga tggtcgtctt gcctgacccg cctttctggt
      421 taagtacagc gataaccttc atgcgttccc cttgcgtatt tgtttattta ctcatcgcat
      481 catatacgca gcgaccgcat gacgcaagct gttttactca aatacacatc acctttttag
      541 acggcggcgc tcggtttctt cagcggccaa gctggccggc caggccgcca gcttggcatc
      601 agacaaaccg gccaggattt catgcagccg cacggctgac tcccgagggg aaccctgtgg
      661 ttggcatgca catacaaatg gacgaacgga taaacctttt cacgcccttt taaatatccg
      721 attattctaa taaacgctct tttctcttag gtttacccgc caatatatcc tgtcaaacac
      781 tgatagttta aacggatcac ttcgtgtccc tcggtcacat gtgcatcctc tagtagcggt
      841 cctgctgagc ctcgacatgt tgtcgcaaaa ttcgccctgg acccgcccaa cgatttgtcg
      901 tcactgtcaa ggtttgacct gcacttcatt tggggcccac atacaccaaa aaaatgctgc
      961 ataattctcg gggcagcaag tcggttaccc ggccgccgtg ctggaccggg ttgaatggtg
     1021 cccgtaactt tcggtagagc ggacggccaa tactcaactt caaggaatct cacccatgcg
     1081 cgccggcggg gaaccggagt tcccttcagt gagcgttatt agttcgccgc tcggtgtgtc
     1141 gtagatacta gcccctgggg cacttttgaa atttgaataa gatttatgta atcagtcttt
     1201 taggtttgac cggttctgcc gcttttttta aaattggatt tgtaataata aaacgcaatt
     1261 gtttgttatt gtggcgctct atcatagatg tcgctataaa cctattcagc acaatatatt
     1321 gttttcattt taatattgta catataagta gtagggtaca atcagtaaat tgaacggaga
     1381 atattattca taaaaatacg atagtaacgg gtgatatatt cattagaatg aaccgaaacc
     1441 ggcggtaagg atctgagcta cacatgctca ggttttttac aacgtgcaca acagaattga
     1501 aagcaaatat catgcgatca taggcttctc gcatatctca ttaaagcagg acaagctcat
     1561 tttccagaaa gcctttgctc aagagtctgt gcgatccagt aattcctgtc gtcctgcttt
     1621 gctgtagcga tcatctttcc caatccagcg tcgtcgatac ctgccttctt gaaccaagac
     1681 acaagcaact cgtaagggtc atttctcttc atcttccaca cgtagtggaa ccagatctca
     1741 aggatagggt tcttcaacaa tgtagtacca gcatcgttca atttaagaag ttggaacact
     1801 tccatctctg taatatggtt cttcctccat gctgttgaca actccttctc caactgtcca
     1861 gagatcctct tgttcacacc tgactgagat gaaacgatca tctttgccac agtcacgtcg
     1921 tcgtattgct ccctcaattt caacaaaaga gcttcgatgt aatctgttga aggtgacttc
     1981 acgtatgaaa gccatgctgg caatcctgga ttgttcaaaa cgttctgacc tccagcgtcc
     2041 aactgcaaga tagcgtacac gtcgtcagct gacttacctg actccttcca agacttctgc
     2101 tggatctcct tcaactgata caccactgtg tccctagaag tcctcttagc agttgccaac
     2161 atcctagcca actcgtcgtc accgtagtga gcagccattg tcttgaagat agccctgtct
     2221 gcaggcaacc tacccctctt gcagatctta gccacgtagt gtgcccagtc tgtgaactca
     2281 gaagaagaaa acacgtcagt tttaacctgc ttaaccctca acacttcaaa cctgttgtta
     2341 accttaagct gcttgtcaag agagaaagtc aacttagcgt tgtgtgccaa gtccttaagt
     2401 gccttcacca cagaaaaaac cattgtatcg ataattgtaa atgtaattgt aatgttgttt
     2461 gttgtttgtt gttgttggta attgttgtaa aaatgagctc ttatactcga gcgtgtcctc
     2521 tccaaatgaa atgaacttcc ttatatagag gaagggtctt gcgaaggata gtgggattgt
     2581 gcgtcatccc ttacgtcagt ggagatgtca catcaatcca cttgctttgt agacgtggtt
     2641 ggaacctctt ctttttccac gatgctcctc gtgggtgggg gtccatcttt gggaccactg
     2701 tcggcagaga gatcttgaat gatagccttt cctttatcgc aatgatggca tttgtaggag
     2761 ccaccttcct tttctactgt cctttcgatg aagtgacaga tagctgggca atggaatccg
     2821 aggaggtttc ccgaaattat cctttgttga aaagtctcaa tagccctttg atcttctgag
     2881 actgtatctt tgacattttt ggagtagacc agagtgtcgt gctccaccat gttgacgaag
     2941 attttcttct tgtcattgag tcgtaaaaga ctctgtatga actgttcgcc agtcttgacg
     3001 gcgagttctg ttagatcctc gatttgaatc ttagactcca tgcatggcct tagattcagt
     3061 aggaactacc tttttagaga ctccaatctc tattacttgc cttggtttat gaagcaagcc
     3121 ttgaatcgtc catactggaa tagtacttct gatcttgaga aatatgtctt tctctgtgtt
     3181 cttgatgcaa ttagtcctga atcttttgac tgcatcttta accttcttgg gaaggtattt
     3241 gatctcctgg agattgttac tcgggtagat cgtcttgatg agacttgctg cgtaggcctc
     3301 tctaaccatc tgtgggtcag cattctttct gaaattgaag aggctaacct tctcattatc
     3361 agtggtgaac atagtgtcgt caccttcacc ttcgaacttc cttcctagat cgtaaagata
     3421 gaggaaatcg tccattgtaa tctccggggc aaaggagatc tcttttgggg ctggatcact
     3481 gctgggcctt ttggttccta gcgtgagcca gtgggctttt tgctttggtg ggcttgttag
     3541 ggccttagca aagctcttgg gcttgagttg agcttctcct ttggggatga agttcaacct
     3601 gtctgtttgc tgacttgttg tgtacgcgtc agctgctgct cttgcctctg taatagtggc
     3661 aaatttcttg tgtgcaactc cgggaacgcc gtttgttgcc gcctttgtac aaccccagtc
     3721 atcgtatata ccggcatgtg gaccgttata cacaacgtag tagttgatat gagggtgttg
     3781 aatacccgat tctgctctga gaggagcaac tgtgctgtta agctcagatt tttgtgggat
     3841 tggaattcct ccaagcttga attcttgcag cgctccaatt cagcaagcgt cgatctagta
     3901 acatagatga caccgcgcgc gataatttat cctagtttgc gcgctatatt ttgttttcta
     3961 tcgcgtatta aatgtataat tgcgggactc taatcataaa aacccatctc ataaataacg
     4021 tcatgcatta catgttaatt attacatgct taacgtaatt caacagaaat tatatgataa
     4081 tcatcgcaag accggcaaca ggattcaatc ttaagaaact ttattgccaa atgtttgaac
     4141 gatctgcttg actctagatc cagagtcccg ctcaaatctc agtcacagga agaacaggtc
     4201 taggaggaac aggaagagag aaatcaagct gccagaatcc cacatcatgc caatttccat
     4261 gtttgaaacc agcagctcta agcattcccc taggagcata accaagagcc tcatgcatcc
     4321 tcacagaagg atcattagga agtccaatca cagcaaccac agacttgaaa ccctgagcct
     4381 ccaaagactt caaaagatga gtgtaaagag tagatccaag accagttctt tgatgcctag
     4441 gagacacgta aacagtagac tcagcagtcc aatcgtaagc attcctagcc ttccaaggac
     4501 cagcataagc aataccagcc acttcaccat ccacttcagc aacaagccaa ggatacctct
     4561 ctctaagcct cacaagatca tcagtccact cttgtggttc ttgaggctca gtcctgaaat
     4621 tcacagtaga agtctcaatg taatgattca caatagtgca cacagcaggc atatcagcct
     4681 cagtagccct tctaatatca gcaggccttc tttcaggaga catgcgaaac gatccagatc
     4741 cggtgcagat tatttggatt gagagtgaat atgagactct aattggatac cgaggggaat
     4801 ttatggaacg tcagtggagc attttgacaa gaaatatttg ctagctgata gtgaccttag
     4861 gcgacttttg aacgcgcaat aatggtttct gacgtatgtg cttagctcat taaactccag
     4921 aaacccgcgg ctgagtggct ccttcaacgt tgcggttctg tcagttccaa acgtaaaacg
     4981 gcttgtcccg cgtcatcggc gggggtcata acgtgactcc cttaattctc cgctccggat
     5041 ccgaattcgg cacagagtgt tcaaccccag tacattaaaa acgtccgcaa tgtgttatta
     5101 agttgtctaa gcgtcaattt gtttacacca caatatatcc tgccaccagc cagccaacag
     5161 ctccccgacc ggcagctcgg cacaaaatca ccactcgata caggcagccc atcaaccatg
     5221 actactgtcc ggtccaacag atccgagcag gaaccgtaaa aaggccgcgt tgctggcgtt
     5281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5881 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5941 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     6001 atcaatctaa agtatatatg agtaaacttg gtctgacagc taaaacaatt catccagtaa
     6061 aatataatat tttattttct cccaatcagg cttgatcccc agtaagtcaa aaaatagctc
     6121 gacatactgt tcttccccga tatcctccct gagcgaccgg acgcagaagg caatgtcata
     6181 ccacttgtcc gccctgccgc ttctcccaag atcaataaag ccacttactt tgccatcttt
     6241 cacaaagatg ttgctgtctc ccaggtcgcc gtgggaaaag acaagttcct cttcgggctt
     6301 ttccgtcttt aaaaaatcat acagctcgcg cggatcttta aatggagtgt cctcttccca
     6361 gttttcgcaa tccacatcgg ccagatcgtt attcagtaag taatccaatt cggctaagcg
     6421 gctgtctaag ctattcgtat agggacaatc cgatatgtcg atggagtgaa agagcctgat
     6481 gcactccgca tacagctcga taatcttttc agggctttgt tcatcttcat actcttccga
     6541 gcaaaggacg ccatcggcct cactcatgag cagattgctc cagccatcat gccgttcaaa
     6601 gtgcaggacc tttggaacag gcagctttcc ttccagccat agcatcatgt ccttttcccg
     6661 ttccacatca taggtggtcc ctttataccg gctgtccgtc atttttaaat ataggttttc
     6721 attttctccc accagcttat ataccttagc aggagacatt ccttccgtat cttttacgca
     6781 gcggtatttt tcgatcagtt ttttcaattc cggtgatatt ctcattttag ccatactctt
     6841 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     6901 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc actcggagcc ttgatgtggg
     6961 cgctttttgc agctgttcgg ctgtgcgctg gccagacagt tatgcacagg ccaggcgggt
     7021 tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa aaatcgcctt ttttctcttt
     7081 tatatcagtc acttacatgt gtgaccggtt cccaatgtac ggctttgggt tcccaatgta
     7141 cgggttccgg ttcccaatgt acggctttgg gttcccaatg tacgtgctat ccacaggaaa
     7201 gaagcttttt cgaccttttt cccctgctag ggcaatttgc cctagcatct gctccgtaca
     7261 ttaggaaccg gcggatgctt cgccctcgat caggttgcgg tagcgcatga ctaggatcgg
     7321 gccagcctgc cccgcctcct ccttcaaatc gtactccggc aggtcatttg acccgatcag
     7381 cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg ctgccactgc gttcgtagat
     7441 cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg cggcgtgcca ggcggtagag
     7501 aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg tgaaccgtca gcacgtccgg
     7561 gttcttgcct tctgtgatct cgcggtacat ccaatcagct agctcgatct cgatgtactc
     7621 cggccgcccg gtttcgctct ttacgatctt gtagcggcta atcaaggctt caccctcgga
     7681 taccgtcacc aggcggccgt tcttggcctt cttcgtacgc tgcatggcaa cgtgcgtggt
     7741 gtttaaccga atgcaggttt ctaccaggtc gtctttctgc tttccgccat cggctcgccg
     7801 gcagaacttg agtacgtccg caacgtgtgg acggaacacg cggccgggct tgtctccctt
     7861 cccttcccgg tatcggttca tggattcggt tagatgggaa accgccatca gtaccaggtc
     7921 gtaatcccac acactggcca tgccggccgg ccctgcggaa acctctacgt gcccgtctgg
     7981 aagctcgtag cggatcacct cgccagctcg tcggtcacgc ttcgacagac ggaaaacggc
     8041 cacgtccatg atgctgcgac tatcgcgggt gcccacgtca tagagcatcg gaacgaaaaa
     8101 atctggttgc tcgtcgccct tgggcggctt cctaatcgac gacgcaccgg ctgccggcgg
     8161 ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc cacgattcac cggggcgtgc
     8221 ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc ttcaacttct ccaccaggtc
     8281 atcacccagc gccgcgccga tttgtaccgg gccggatggt ttgcgaccgc tcacgccgat
     8341 tcctcgggct tgggggttcc agtgccattg cagggccggc agacaaccca gccgcttacg
     8401 cctggccaac cgcccgttcc tccacacatg gggcattcca cggcgtcggt gcctggttgt
     8461 tcttgatttt ccatgccgcc tcctttagcc gctaaaattc atctactcat ttattcattt
     8521 gctcatttac tctggtagct gcgcgatgta ttcagatagc agctcggtaa tggtcttgcc
     8581 ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc gccggcaact gaaagttgac
     8641 ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt gcagccttgc tgctgcgtgc
     8701 gctcggacgg ccggcactta gcgtgtttgt gcttttgctc attttctctt tacctcatta
     8761 actcaaatga gttttgattt aatttcagcg gccagcgcct ggacctcgcg ggcagcgtcg
     8821 ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg cagtgcctgg gtagctcacg
     8881 cgctgcgtga tacgggactc aagaatgggc agctcgtacc cggccagcgc ctcggcaacc
     8941 tcaccgccga
//
