LOCUS       sequence-342111-        8581 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 8581)
  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 8581)
  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_00582
FEATURES             Location/Qualifiers
     source          1..8581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    720..744
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(811..1518)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(2114..2459)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2151..2171)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3492..3744)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4344..4526)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4380..4558)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4721..4745
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4881..5469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4961..4980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5640..6434)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6435..6539)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6601..6795
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6861..7934)
                     /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(8363..8581,1..411))
                     /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 aaaggccgct tgtagccttc catccgtgac ctcaatgcgc tgcttaacca gctccaccag
       61 gtcggcggtg gcccatatgt cgtaagggct tggctgcacc ggaatcagca cgaagtcggc
      121 tgccttgatc gcggacacag ccaagtccgc cgcctggggc gctccgtcga tcactacgaa
      181 gtcgcgccgg ccgatggcct tcacgtcgcg gtcaatcgtc gggcggtcga tgccgacaac
      241 ggttagcggt tgatcttccc gcacggccgc ccaatcgcgg gcactgccct ggggatcgga
      301 atcgactaac agaacatcgg ccccggcgag ttgcagggcg cgggctagat gggttgcgat
      361 ggtcgtcttg cctgacccgc ctttctggtt aagtacagcg ataaccttca tgcgttcccc
      421 ttgcgtattt gtttatttac tcatcgcatc atatacgcag cgaccgcatg acgcaagctg
      481 ttttactcaa atacacatca cctttttaga cggcggcgct cggtttcttc agcggccaag
      541 ctggccggcc aggccgccag cttggcatca gacaaaccgg ccaggatttc atgcagccgc
      601 acggctgact cccgagggga accctgtggt tggcatgcac atacaaatgg acgaacggat
      661 aaaccttttc acgccctttt aaatatccga ttattctaat aaacgctctt ttctcttagg
      721 tttacccgcc aatatatcct gtcaaacact gatagtttaa acggatcact tcgtgtccct
      781 cggtcacatg tgcatcctct agtagcggtc ctgctgagcc tcgacatgtt gtcgcaaaat
      841 tcgccctgga cccgcccaac gatttgtcgt cactgtcaag gtttgacctg cacttcattt
      901 ggggcccaca tacaccaaaa aaatgctgca taattctcgg ggcagcaagt cggttacccg
      961 gccgccgtgc tggaccgggt tgaatggtgc ccgtaacttt cggtagagcg gacggccaat
     1021 actcaacttc aaggaatctc acccatgcgc gccggcgggg aaccggagtt cccttcagtg
     1081 agcgttatta gttcgccgct cggtgtgtcg tagatactag cccctggggc acttttgaaa
     1141 tttgaataag atttatgtaa tcagtctttt aggtttgacc ggttctgccg ctttttttaa
     1201 aattggattt gtaataataa aacgcaattg tttgttattg tggcgctcta tcatagatgt
     1261 cgctataaac ctattcagca caatatattg ttttcatttt aatattgtac atataagtag
     1321 tagggtacaa tcagtaaatt gaacggagaa tattattcat aaaaatacga tagtaacggg
     1381 tgatatattc attagaatga accgaaaccg gcggtaagga tctgagctac acatgctcag
     1441 gttttttaca acgtgcacaa cagaattgaa agcaaatatc atgcgatcat aggcttctcg
     1501 catatctcat taaagcagga caagctcaag cgtcaagctt ctcaagcaac tttgggtacc
     1561 tagcgtagta gatagtcctg tagttcttgt agtgcttgta gttcactgaa gtagtaggat
     1621 agattccctt cttagccaac tccttgaact tgaacttttt gtacacgtcg tcaggagtca
     1681 tctctgccat caaccacctg ttgtacatct tgtcgtcctt agcagcctgt ttagccaact
     1741 tctccaactc gttctgctgc aaagccttaa gctgctgagc agccttctcg ttctttgcct
     1801 tcaagttctg ctcagccttg atagcctggt ccttcacctt ctgctctgcc ttcactgcct
     1861 ttgcctgcaa cttagcctcg atcttagaca atttttcttt agtgtttgac atctccttag
     1921 cctgcttagc aaccaacttc tccttaggag tgatctttga cctagctgcc cagttcaact
     1981 tcataaatga caactcagac ttcaaagcgt ccactgctcc cattgtatcg ataattgtaa
     2041 atgtaattgt aatgttgttt gttgtttgtt gttgttggta attgttgtaa aaatgagctc
     2101 ttatactcga gcgtgtcctc tccaaatgaa atgaacttcc ttatatagag gaagggtctt
     2161 gcgaaggata gtgggattgt gcgtcatccc ttacgtcagt ggagatgtca catcaatcca
     2221 cttgctttgt agacgtggtt ggaacctctt ctttttccac gatgctcctc gtgggtgggg
     2281 gtccatcttt gggaccactg tcggcagaga gatcttgaat gatagccttt cctttatcgc
     2341 aatgatggca tttgtaggag ccaccttcct tttctactgt cctttcgatg aagtgacaga
     2401 tagctgggca atggaatccg aggaggtttc ccgaaattat cctttgttga aaagtctcaa
     2461 tagccctttg atcttctgag actgtatctt tgacattttt ggagtagacc agagtgtcgt
     2521 gctccaccat gttgacgaag attttcttct tgtcattgag tcgtaaaaga ctctgtatga
     2581 actgttcgcc agtcttgacg gcgagttctg ttagatcctc gatttgaatc ttagactcca
     2641 tgcatggcct tagattcagt aggaactacc tttttagaga ctccaatctc tattacttgc
     2701 cttggtttat gaagcaagcc ttgaatcgtc catactggaa tagtacttct gatcttgaga
     2761 aatatgtctt tctctgtgtt cttgatgcaa ttagtcctga atcttttgac tgcatcttta
     2821 accttcttgg gaaggtattt gatctcctgg agattgttac tcgggtagat cgtcttgatg
     2881 agacttgctg cgtaggcctc tctaaccatc tgtgggtcag cattctttct gaaattgaag
     2941 aggctaacct tctcattatc agtggtgaac atagtgtcgt caccttcacc ttcgaacttc
     3001 cttcctagat cgtaaagata gaggaaatcg tccattgtaa tctccggggc aaaggagatc
     3061 tcttttgggg ctggatcact gctgggcctt ttggttccta gcgtgagcca gtgggctttt
     3121 tgctttggtg ggcttgttag ggccttagca aagctcttgg gcttgagttg agcttctcct
     3181 ttggggatga agttcaacct gtctgtttgc tgacttgttg tgtacgcgtc agctgctgct
     3241 cttgcctctg taatagtggc aaatttcttg tgtgcaactc cgggaacgcc gtttgttgcc
     3301 gcctttgtac aaccccagtc atcgtatata ccggcatgtg gaccgttata cacaacgtag
     3361 tagttgatat gagggtgttg aatacccgat tctgctctga gaggagcaac tgtgctgtta
     3421 agctcagatt tttgtgggat tggaattcct ccaagcttga attcttgcag cgctccaatt
     3481 cagcaagcgt cgatctagta acatagatga caccgcgcgc gataatttat cctagtttgc
     3541 gcgctatatt ttgttttcta tcgcgtatta aatgtataat tgcgggactc taatcataaa
     3601 aacccatctc ataaataacg tcatgcatta catgttaatt attacatgct taacgtaatt
     3661 caacagaaat tatatgataa tcatcgcaag accggcaaca ggattcaatc ttaagaaact
     3721 ttattgccaa atgtttgaac gatctgcttg actctagatc cagagtcccg ctcaaatctc
     3781 agtcacagga agaacaggtc taggaggaac aggaagagag aaatcaagct gccagaatcc
     3841 cacatcatgc caatttccat gtttgaaacc agcagctcta agcattcccc taggagcata
     3901 accaagagcc tcatgcatcc tcacagaagg atcattagga agtccaatca cagcaaccac
     3961 agacttgaaa ccctgagcct ccaaagactt caaaagatga gtgtaaagag tagatccaag
     4021 accagttctt tgatgcctag gagacacgta aacagtagac tcagcagtcc aatcgtaagc
     4081 attcctagcc ttccaaggac cagcataagc aataccagcc acttcaccat ccacttcagc
     4141 aacaagccaa ggatacctct ctctaagcct cacaagatca tcagtccact cttgtggttc
     4201 ttgaggctca gtcctgaaat tcacagtaga agtctcaatg taatgattca caatagtgca
     4261 cacagcaggc atatcagcct cagtagccct tctaatatca gcaggccttc tttcaggaga
     4321 catgcgaaac gatccagatc cggtgcagat tatttggatt gagagtgaat atgagactct
     4381 aattggatac cgaggggaat ttatggaacg tcagtggagc attttgacaa gaaatatttg
     4441 ctagctgata gtgaccttag gcgacttttg aacgcgcaat aatggtttct gacgtatgtg
     4501 cttagctcat taaactccag aaacccgcgg ctgagtggct ccttcaacgt tgcggttctg
     4561 tcagttccaa acgtaaaacg gcttgtcccg cgtcatcggc gggggtcata acgtgactcc
     4621 cttaattctc cgctccggat ccgaattcgg cacagagtgt tcaaccccag tacattaaaa
     4681 acgtccgcaa tgtgttatta agttgtctaa gcgtcaattt gtttacacca caatatatcc
     4741 tgccaccagc cagccaacag ctccccgacc ggcagctcgg cacaaaatca ccactcgata
     4801 caggcagccc atcaaccatg actactgtcc ggtccaacag atccgagcag gaaccgtaaa
     4861 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     4921 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     4981 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5041 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5101 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5161 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5221 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5281 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5341 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     5401 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     5461 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     5521 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     5581 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagc
     5641 taaaacaatt catccagtaa aatataatat tttattttct cccaatcagg cttgatcccc
     5701 agtaagtcaa aaaatagctc gacatactgt tcttccccga tatcctccct gagcgaccgg
     5761 acgcagaagg caatgtcata ccacttgtcc gccctgccgc ttctcccaag atcaataaag
     5821 ccacttactt tgccatcttt cacaaagatg ttgctgtctc ccaggtcgcc gtgggaaaag
     5881 acaagttcct cttcgggctt ttccgtcttt aaaaaatcat acagctcgcg cggatcttta
     5941 aatggagtgt cctcttccca gttttcgcaa tccacatcgg ccagatcgtt attcagtaag
     6001 taatccaatt cggctaagcg gctgtctaag ctattcgtat agggacaatc cgatatgtcg
     6061 atggagtgaa agagcctgat gcactccgca tacagctcga taatcttttc agggctttgt
     6121 tcatcttcat actcttccga gcaaaggacg ccatcggcct cactcatgag cagattgctc
     6181 cagccatcat gccgttcaaa gtgcaggacc tttggaacag gcagctttcc ttccagccat
     6241 agcatcatgt ccttttcccg ttccacatca taggtggtcc ctttataccg gctgtccgtc
     6301 atttttaaat ataggttttc attttctccc accagcttat ataccttagc aggagacatt
     6361 ccttccgtat cttttacgca gcggtatttt tcgatcagtt ttttcaattc cggtgatatt
     6421 ctcattttag ccatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6481 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6541 actcggagcc ttgatgtggg cgctttttgc agctgttcgg ctgtgcgctg gccagacagt
     6601 tatgcacagg ccaggcgggt tttaagagtt ttaataagtt ttaaagagtt ttaggcggaa
     6661 aaatcgcctt ttttctcttt tatatcagtc acttacatgt gtgaccggtt cccaatgtac
     6721 ggctttgggt tcccaatgta cgggttccgg ttcccaatgt acggctttgg gttcccaatg
     6781 tacgtgctat ccacaggaaa gaagcttttt cgaccttttt cccctgctag ggcaatttgc
     6841 cctagcatct gctccgtaca ttaggaaccg gcggatgctt cgccctcgat caggttgcgg
     6901 tagcgcatga ctaggatcgg gccagcctgc cccgcctcct ccttcaaatc gtactccggc
     6961 aggtcatttg acccgatcag cttgcgcacg gtgaaacaga acttcttgaa ctctccggcg
     7021 ctgccactgc gttcgtagat cgtcttgaac aaccatctgg cttctgcctt gcctgcggcg
     7081 cggcgtgcca ggcggtagag aaaacggccg atgccgggat cgatcaaaaa gtaatcgggg
     7141 tgaaccgtca gcacgtccgg gttcttgcct tctgtgatct cgcggtacat ccaatcagct
     7201 agctcgatct cgatgtactc cggccgcccg gtttcgctct ttacgatctt gtagcggcta
     7261 atcaaggctt caccctcgga taccgtcacc aggcggccgt tcttggcctt cttcgtacgc
     7321 tgcatggcaa cgtgcgtggt gtttaaccga atgcaggttt ctaccaggtc gtctttctgc
     7381 tttccgccat cggctcgccg gcagaacttg agtacgtccg caacgtgtgg acggaacacg
     7441 cggccgggct tgtctccctt cccttcccgg tatcggttca tggattcggt tagatgggaa
     7501 accgccatca gtaccaggtc gtaatcccac acactggcca tgccggccgg ccctgcggaa
     7561 acctctacgt gcccgtctgg aagctcgtag cggatcacct cgccagctcg tcggtcacgc
     7621 ttcgacagac ggaaaacggc cacgtccatg atgctgcgac tatcgcgggt gcccacgtca
     7681 tagagcatcg gaacgaaaaa atctggttgc tcgtcgccct tgggcggctt cctaatcgac
     7741 gacgcaccgg ctgccggcgg ttgccgggat tctttgcgga ttcgatcagc ggccgcttgc
     7801 cacgattcac cggggcgtgc ttctgcctcg atgcgttgcc gctgggcggc ctgcgcggcc
     7861 ttcaacttct ccaccaggtc atcacccagc gccgcgccga tttgtaccgg gccggatggt
     7921 ttgcgaccgc tcacgccgat tcctcgggct tgggggttcc agtgccattg cagggccggc
     7981 agacaaccca gccgcttacg cctggccaac cgcccgttcc tccacacatg gggcattcca
     8041 cggcgtcggt gcctggttgt tcttgatttt ccatgccgcc tcctttagcc gctaaaattc
     8101 atctactcat ttattcattt gctcatttac tctggtagct gcgcgatgta ttcagatagc
     8161 agctcggtaa tggtcttgcc ttggcgtacc gcgtacatct tcagcttggt gtgatcctcc
     8221 gccggcaact gaaagttgac ccgcttcatg gctggcgtgt ctgccaggct ggccaacgtt
     8281 gcagccttgc tgctgcgtgc gctcggacgg ccggcactta gcgtgtttgt gcttttgctc
     8341 attttctctt tacctcatta actcaaatga gttttgattt aatttcagcg gccagcgcct
     8401 ggacctcgcg ggcagcgtcg ccctcgggtt ctgattcaag aacggttgtg ccggcggcgg
     8461 cagtgcctgg gtagctcacg cgctgcgtga tacgggactc aagaatgggc agctcgtacc
     8521 cggccagcgc ctcggcaacc tcaccgccga tgcgcgtgcc tttgatcgcc cgcgacacga
     8581 c
//
