LOCUS       sequence-342549-        8389 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 8389)
  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 8389)
  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_15753
FEATURES             Location/Qualifiers
     source          1..8389
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(214..843)
                     /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    1152..1176
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1243..1950)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2354..2699)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2391..2411)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3732..3984)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4584..4766)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4620..4798)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4961..4985
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(5121..5709)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5201..5220)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5880..6674)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6675..6779)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6841..7035
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(7101..8174)
                     /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 attcagatag cagctcggta atggtcttgc cttggcgtac cgcgtacatc ttcagcttgg
       61 tgtgatcctc cgccggcaac tgaaagttga cccgcttcat ggctggcgtg tctgccaggc
      121 tggccaacgt tgcagccttg ctgctgcgtg cgctcggacg gccggcactt agcgtgtttg
      181 tgcttttgct cattttctct ttacctcatt aactcaaatg agttttgatt taatttcagc
      241 ggccagcgcc tggacctcgc gggcagcgtc gccctcgggt tctgattcaa gaacggttgt
      301 gccggcggcg gcagtgcctg ggtagctcac gcgctgcgtg atacgggact caagaatggg
      361 cagctcgtac ccggccagcg cctcggcaac ctcaccgccg atgcgcgtgc ctttgatcgc
      421 ccgcgacacg acaaaggccg cttgtagcct tccatccgtg acctcaatgc gctgcttaac
      481 cagctccacc aggtcggcgg tggcccatat gtcgtaaggg cttggctgca ccggaatcag
      541 cacgaagtcg gctgccttga tcgcggacac agccaagtcc gccgcctggg gcgctccgtc
      601 gatcactacg aagtcgcgcc ggccgatggc cttcacgtcg cggtcaatcg tcgggcggtc
      661 gatgccgaca acggttagcg gttgatcttc ccgcacggcc gcccaatcgc gggcactgcc
      721 ctggggatcg gaatcgacta acagaacatc ggccccggcg agttgcaggg cgcgggctag
      781 atgggttgcg atggtcgtct tgcctgaccc gcctttctgg ttaagtacag cgataacctt
      841 catgcgttcc ccttgcgtat ttgtttattt actcatcgca tcatatacgc agcgaccgca
      901 tgacgcaagc tgttttactc aaatacacat caccttttta gacggcggcg ctcggtttct
      961 tcagcggcca agctggccgg ccaggccgcc agcttggcat cagacaaacc ggccaggatt
     1021 tcatgcagcc gcacggctga ctcccgaggg gaaccctgtg gttggcatgc acatacaaat
     1081 ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc gattattcta ataaacgctc
     1141 ttttctctta ggtttacccg ccaatatatc ctgtcaaaca ctgatagttt aaacggatca
     1201 cttcgtgtcc ctcggtcaca tgtgcatcct ctagtagcgg tcctgctgag cctcgacatg
     1261 ttgtcgcaaa attcgccctg gacccgccca acgatttgtc gtcactgtca aggtttgacc
     1321 tgcacttcat ttggggccca catacaccaa aaaaatgctg cataattctc ggggcagcaa
     1381 gtcggttacc cggccgccgt gctggaccgg gttgaatggt gcccgtaact ttcggtagag
     1441 cggacggcca atactcaact tcaaggaatc tcacccatgc gcgccggcgg ggaaccggag
     1501 ttcccttcag tgagcgttat tagttcgccg ctcggtgtgt cgtagatact agcccctggg
     1561 gcacttttga aatttgaata agatttatgt aatcagtctt ttaggtttga ccggttctgc
     1621 cgcttttttt aaaattggat ttgtaataat aaaacgcaat tgtttgttat tgtggcgctc
     1681 tatcatagat gtcgctataa acctattcag cacaatatat tgttttcatt ttaatattgt
     1741 acatataagt agtagggtac aatcagtaaa ttgaacggag aatattattc ataaaaatac
     1801 gatagtaacg ggtgatatat tcattagaat gaaccgaaac cggcggtaag gatctgagct
     1861 acacatgctc aggtttttta caacgtgcac aacagaattg aaagcaaata tcatgcgatc
     1921 ataggcttct cgcatatctc attaaagcag gacaagctca cacagccaca gcgaaagtgt
     1981 tcacccaagc tgggttcttc accttgtaag cagcagcgta cttgtaccac aagttgaact
     2041 caggagaagg gtccaatccc acgttcctgt ttccgttcct acccaacttc aacctcttgt
     2101 acacgatgtt agggtccact ccctgcttca acaacttctg cacctcagtg tccaagtcca
     2161 tcttagcagc gtagaaccta agcttagtag aaagcttctt gtctccgaac cacttagaca
     2221 ccctagatcc gaagttcttg aacacagatg ggatcacagc cattgtatcg ataattgtaa
     2281 atgtaattgt aatgttgttt gttgtttgtt gttgttggta attgttgtaa aaatgagctc
     2341 ttatactcga gcgtgtcctc tccaaatgaa atgaacttcc ttatatagag gaagggtctt
     2401 gcgaaggata gtgggattgt gcgtcatccc ttacgtcagt ggagatgtca catcaatcca
     2461 cttgctttgt agacgtggtt ggaacctctt ctttttccac gatgctcctc gtgggtgggg
     2521 gtccatcttt gggaccactg tcggcagaga gatcttgaat gatagccttt cctttatcgc
     2581 aatgatggca tttgtaggag ccaccttcct tttctactgt cctttcgatg aagtgacaga
     2641 tagctgggca atggaatccg aggaggtttc ccgaaattat cctttgttga aaagtctcaa
     2701 tagccctttg atcttctgag actgtatctt tgacattttt ggagtagacc agagtgtcgt
     2761 gctccaccat gttgacgaag attttcttct tgtcattgag tcgtaaaaga ctctgtatga
     2821 actgttcgcc agtcttgacg gcgagttctg ttagatcctc gatttgaatc ttagactcca
     2881 tgcatggcct tagattcagt aggaactacc tttttagaga ctccaatctc tattacttgc
     2941 cttggtttat gaagcaagcc ttgaatcgtc catactggaa tagtacttct gatcttgaga
     3001 aatatgtctt tctctgtgtt cttgatgcaa ttagtcctga atcttttgac tgcatcttta
     3061 accttcttgg gaaggtattt gatctcctgg agattgttac tcgggtagat cgtcttgatg
     3121 agacttgctg cgtaggcctc tctaaccatc tgtgggtcag cattctttct gaaattgaag
     3181 aggctaacct tctcattatc agtggtgaac atagtgtcgt caccttcacc ttcgaacttc
     3241 cttcctagat cgtaaagata gaggaaatcg tccattgtaa tctccggggc aaaggagatc
     3301 tcttttgggg ctggatcact gctgggcctt ttggttccta gcgtgagcca gtgggctttt
     3361 tgctttggtg ggcttgttag ggccttagca aagctcttgg gcttgagttg agcttctcct
     3421 ttggggatga agttcaacct gtctgtttgc tgacttgttg tgtacgcgtc agctgctgct
     3481 cttgcctctg taatagtggc aaatttcttg tgtgcaactc cgggaacgcc gtttgttgcc
     3541 gcctttgtac aaccccagtc atcgtatata ccggcatgtg gaccgttata cacaacgtag
     3601 tagttgatat gagggtgttg aatacccgat tctgctctga gaggagcaac tgtgctgtta
     3661 agctcagatt tttgtgggat tggaattcct ccaagcttga attcttgcag cgctccaatt
     3721 cagcaagcgt cgatctagta acatagatga caccgcgcgc gataatttat cctagtttgc
     3781 gcgctatatt ttgttttcta tcgcgtatta aatgtataat tgcgggactc taatcataaa
     3841 aacccatctc ataaataacg tcatgcatta catgttaatt attacatgct taacgtaatt
     3901 caacagaaat tatatgataa tcatcgcaag accggcaaca ggattcaatc ttaagaaact
     3961 ttattgccaa atgtttgaac gatctgcttg actctagatc cagagtcccg ctcaaatctc
     4021 agtcacagga agaacaggtc taggaggaac aggaagagag aaatcaagct gccagaatcc
     4081 cacatcatgc caatttccat gtttgaaacc agcagctcta agcattcccc taggagcata
     4141 accaagagcc tcatgcatcc tcacagaagg atcattagga agtccaatca cagcaaccac
     4201 agacttgaaa ccctgagcct ccaaagactt caaaagatga gtgtaaagag tagatccaag
     4261 accagttctt tgatgcctag gagacacgta aacagtagac tcagcagtcc aatcgtaagc
     4321 attcctagcc ttccaaggac cagcataagc aataccagcc acttcaccat ccacttcagc
     4381 aacaagccaa ggatacctct ctctaagcct cacaagatca tcagtccact cttgtggttc
     4441 ttgaggctca gtcctgaaat tcacagtaga agtctcaatg taatgattca caatagtgca
     4501 cacagcaggc atatcagcct cagtagccct tctaatatca gcaggccttc tttcaggaga
     4561 catgcgaaac gatccagatc cggtgcagat tatttggatt gagagtgaat atgagactct
     4621 aattggatac cgaggggaat ttatggaacg tcagtggagc attttgacaa gaaatatttg
     4681 ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct gacgtatgtg
     4741 cttagctcat taaactccag aaacccgcgg ctgagtggct ccttcaacgt tgcggttctg
     4801 tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata acgtgactcc
     4861 cttaattctc cgctccggat ccgaattcgg cacagagtgt tcaaccccag tacattaaaa
     4921 acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc
     4981 tgccaccagc cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata
     5041 caggcagccc atcaaccatg actactgtcc ggtccaacag atccgagcag gaaccgtaaa
     5101 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5161 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5221 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5281 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5341 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5401 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5461 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5521 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5581 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5641 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5701 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5761 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5821 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc
     5881 taaaacaatt catccagtaa aatataatat tttattttct cccaatcagg cttgatcccc
     5941 agtaagtcaa aaaatagctc gacatactgt tcttccccga tatcctccct gagcgaccgg
     6001 acgcagaagg caatgtcata ccacttgtcc gccctgccgc ttctcccaag atcaataaag
     6061 ccacttactt tgccatcttt cacaaagatg ttgctgtctc ccaggtcgcc gtgggaaaag
     6121 acaagttcct cttcgggctt ttccgtcttt aaaaaatcat acagctcgcg cggatcttta
     6181 aatggagtgt cctcttccca gttttcgcaa tccacatcgg ccagatcgtt attcagtaag
     6241 taatccaatt cggctaagcg gctgtctaag ctattcgtat agggacaatc cgatatgtcg
     6301 atggagtgaa agagcctgat gcactccgca tacagctcga taatcttttc agggctttgt
     6361 tcatcttcat actcttccga gcaaaggacg ccatcggcct cactcatgag cagattgctc
     6421 cagccatcat gccgttcaaa gtgcaggacc tttggaacag gcagctttcc ttccagccat
     6481 agcatcatgt ccttttcccg ttccacatca taggtggtcc ctttataccg gctgtccgtc
     6541 atttttaaat ataggttttc attttctccc accagcttat ataccttagc aggagacatt
     6601 ccttccgtat cttttacgca gcggtatttt tcgatcagtt ttttcaattc cggtgatatt
     6661 ctcattttag ccatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6721 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6781 actcggagcc ttgatgtggg cgctttttgc agctgttcgg ctgtgcgctg gccagacagt
     6841 tatgcacagg ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa
     6901 aaatcgcctt ttttctcttt tatatcagtc acttacatgt gtgaccggtt cccaatgtac
     6961 ggctttgggt tcccaatgta cgggttccgg ttcccaatgt acggctttgg gttcccaatg
     7021 tacgtgctat ccacaggaaa gaagcttttt cgaccttttt cccctgctag ggcaatttgc
     7081 cctagcatct gctccgtaca ttaggaaccg gcggatgctt cgccctcgat caggttgcgg
     7141 tagcgcatga ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc gtactccggc
     7201 aggtcatttg acccgatcag cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg
     7261 ctgccactgc gttcgtagat cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg
     7321 cggcgtgcca ggcggtagag aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg
     7381 tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct cgcggtacat ccaatcagct
     7441 agctcgatct cgatgtactc cggccgcccg gtttcgctct ttacgatctt gtagcggcta
     7501 atcaaggctt caccctcgga taccgtcacc aggcggccgt tcttggcctt cttcgtacgc
     7561 tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc gtctttctgc
     7621 tttccgccat cggctcgccg gcagaacttg agtacgtccg caacgtgtgg acggaacacg
     7681 cggccgggct tgtctccctt cccttcccgg tatcggttca tggattcggt tagatgggaa
     7741 accgccatca gtaccaggtc gtaatcccac acactggcca tgccggccgg ccctgcggaa
     7801 acctctacgt gcccgtctgg aagctcgtag cggatcacct cgccagctcg tcggtcacgc
     7861 ttcgacagac ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt gcccacgtca
     7921 tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt cctaatcgac
     7981 gacgcaccgg ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc
     8041 cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc
     8101 ttcaacttct ccaccaggtc atcacccagc gccgcgccga tttgtaccgg gccggatggt
     8161 ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc agtgccattg cagggccggc
     8221 agacaaccca gccgcttacg cctggccaac cgcccgttcc tccacacatg gggcattcca
     8281 cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc tcctttagcc gctaaaattc
     8341 atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgt
//
