LOCUS       sequence-342796-        8242 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 8242)
  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 8242)
  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_22870
FEATURES             Location/Qualifiers
     source          1..8242
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(172..801)
                     /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    1110..1134
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(1201..1908)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     RBS             1992..2000
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(2165..2510)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(2202..2222)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3543..3795)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4395..4577)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4431..4609)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4772..4796
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4932..5520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5012..5031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5691..6485)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6486..6590)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6652..6846
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6912..7985)
                     /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 cgtacatctt cagcttggtg tgatcctccg ccggcaactg aaagttgacc cgcttcatgg
       61 ctggcgtgtc tgccaggctg gccaacgttg cagccttgct gctgcgtgcg ctcggacggc
      121 cggcacttag cgtgtttgtg cttttgctca ttttctcttt acctcattaa ctcaaatgag
      181 ttttgattta atttcagcgg ccagcgcctg gacctcgcgg gcagcgtcgc cctcgggttc
      241 tgattcaaga acggttgtgc cggcggcggc agtgcctggg tagctcacgc gctgcgtgat
      301 acgggactca agaatgggca gctcgtaccc ggccagcgcc tcggcaacct caccgccgat
      361 gcgcgtgcct ttgatcgccc gcgacacgac aaaggccgct tgtagccttc catccgtgac
      421 ctcaatgcgc tgcttaacca gctccaccag gtcggcggtg gcccatatgt cgtaagggct
      481 tggctgcacc ggaatcagca cgaagtcggc tgccttgatc gcggacacag ccaagtccgc
      541 cgcctggggc gctccgtcga tcactacgaa gtcgcgccgg ccgatggcct tcacgtcgcg
      601 gtcaatcgtc gggcggtcga tgccgacaac ggttagcggt tgatcttccc gcacggccgc
      661 ccaatcgcgg gcactgccct ggggatcgga atcgactaac agaacatcgg ccccggcgag
      721 ttgcagggcg cgggctagat gggttgcgat ggtcgtcttg cctgacccgc ctttctggtt
      781 aagtacagcg ataaccttca tgcgttcccc ttgcgtattt gtttatttac tcatcgcatc
      841 atatacgcag cgaccgcatg acgcaagctg ttttactcaa atacacatca cctttttaga
      901 cggcggcgct cggtttcttc agcggccaag ctggccggcc aggccgccag cttggcatca
      961 gacaaaccgg ccaggatttc atgcagccgc acggctgact cccgagggga accctgtggt
     1021 tggcatgcac atacaaatgg acgaacggat aaaccttttc acgccctttt aaatatccga
     1081 ttattctaat aaacgctctt ttctcttagg tttacccgcc aatatatcct gtcaaacact
     1141 gatagtttaa acggatcact tcgtgtccct cggtcacatg tgcatcctct agtagcggtc
     1201 ctgctgagcc tcgacatgtt gtcgcaaaat tcgccctgga cccgcccaac gatttgtcgt
     1261 cactgtcaag gtttgacctg cacttcattt ggggcccaca tacaccaaaa aaatgctgca
     1321 taattctcgg ggcagcaagt cggttacccg gccgccgtgc tggaccgggt tgaatggtgc
     1381 ccgtaacttt cggtagagcg gacggccaat actcaacttc aaggaatctc acccatgcgc
     1441 gccggcgggg aaccggagtt cccttcagtg agcgttatta gttcgccgct cggtgtgtcg
     1501 tagatactag cccctggggc acttttgaaa tttgaataag atttatgtaa tcagtctttt
     1561 aggtttgacc ggttctgccg ctttttttaa aattggattt gtaataataa aacgcaattg
     1621 tttgttattg tggcgctcta tcatagatgt cgctataaac ctattcagca caatatattg
     1681 ttttcatttt aatattgtac atataagtag tagggtacaa tcagtaaatt gaacggagaa
     1741 tattattcat aaaaatacga tagtaacggg tgatatattc attagaatga accgaaaccg
     1801 gcggtaagga tctgagctac acatgctcag gttttttaca acgtgcacaa cagaattgaa
     1861 agcaaatatc atgcgatcat aggcttctcg catatctcat taaagcagga caagctcaag
     1921 acctcttgtc tcccttgatc ttcaagtgct ccaccaactt cttccagtca gcgtcagtga
     1981 tcttctcagt cacctcctta gccttgtcct tagccttcac agcagccttg atagggttca
     2041 acttcttcaa agtgtccttc aaagagaatc ccattgtatc gataattgta aatgtaattg
     2101 taatgttgtt tgttgtttgt tgttgttggt aattgttgta aaaatgagct cttatactcg
     2161 agcgtgtcct ctccaaatga aatgaacttc cttatataga ggaagggtct tgcgaaggat
     2221 agtgggattg tgcgtcatcc cttacgtcag tggagatgtc acatcaatcc acttgctttg
     2281 tagacgtggt tggaacctct tctttttcca cgatgctcct cgtgggtggg ggtccatctt
     2341 tgggaccact gtcggcagag agatcttgaa tgatagcctt tcctttatcg caatgatggc
     2401 atttgtagga gccaccttcc ttttctactg tcctttcgat gaagtgacag atagctgggc
     2461 aatggaatcc gaggaggttt cccgaaatta tcctttgttg aaaagtctca atagcccttt
     2521 gatcttctga gactgtatct ttgacatttt tggagtagac cagagtgtcg tgctccacca
     2581 tgttgacgaa gattttcttc ttgtcattga gtcgtaaaag actctgtatg aactgttcgc
     2641 cagtcttgac ggcgagttct gttagatcct cgatttgaat cttagactcc atgcatggcc
     2701 ttagattcag taggaactac ctttttagag actccaatct ctattacttg ccttggttta
     2761 tgaagcaagc cttgaatcgt ccatactgga atagtacttc tgatcttgag aaatatgtct
     2821 ttctctgtgt tcttgatgca attagtcctg aatcttttga ctgcatcttt aaccttcttg
     2881 ggaaggtatt tgatctcctg gagattgtta ctcgggtaga tcgtcttgat gagacttgct
     2941 gcgtaggcct ctctaaccat ctgtgggtca gcattctttc tgaaattgaa gaggctaacc
     3001 ttctcattat cagtggtgaa catagtgtcg tcaccttcac cttcgaactt ccttcctaga
     3061 tcgtaaagat agaggaaatc gtccattgta atctccgggg caaaggagat ctcttttggg
     3121 gctggatcac tgctgggcct tttggttcct agcgtgagcc agtgggcttt ttgctttggt
     3181 gggcttgtta gggccttagc aaagctcttg ggcttgagtt gagcttctcc tttggggatg
     3241 aagttcaacc tgtctgtttg ctgacttgtt gtgtacgcgt cagctgctgc tcttgcctct
     3301 gtaatagtgg caaatttctt gtgtgcaact ccgggaacgc cgtttgttgc cgcctttgta
     3361 caaccccagt catcgtatat accggcatgt ggaccgttat acacaacgta gtagttgata
     3421 tgagggtgtt gaatacccga ttctgctctg agaggagcaa ctgtgctgtt aagctcagat
     3481 ttttgtggga ttggaattcc tccaagcttg aattcttgca gcgctccaat tcagcaagcg
     3541 tcgatctagt aacatagatg acaccgcgcg cgataattta tcctagtttg cgcgctatat
     3601 tttgttttct atcgcgtatt aaatgtataa ttgcgggact ctaatcataa aaacccatct
     3661 cataaataac gtcatgcatt acatgttaat tattacatgc ttaacgtaat tcaacagaaa
     3721 ttatatgata atcatcgcaa gaccggcaac aggattcaat cttaagaaac tttattgcca
     3781 aatgtttgaa cgatctgctt gactctagat ccagagtccc gctcaaatct cagtcacagg
     3841 aagaacaggt ctaggaggaa caggaagaga gaaatcaagc tgccagaatc ccacatcatg
     3901 ccaatttcca tgtttgaaac cagcagctct aagcattccc ctaggagcat aaccaagagc
     3961 ctcatgcatc ctcacagaag gatcattagg aagtccaatc acagcaacca cagacttgaa
     4021 accctgagcc tccaaagact tcaaaagatg agtgtaaaga gtagatccaa gaccagttct
     4081 ttgatgccta ggagacacgt aaacagtaga ctcagcagtc caatcgtaag cattcctagc
     4141 cttccaagga ccagcataag caataccagc cacttcacca tccacttcag caacaagcca
     4201 aggatacctc tctctaagcc tcacaagatc atcagtccac tcttgtggtt cttgaggctc
     4261 agtcctgaaa ttcacagtag aagtctcaat gtaatgattc acaatagtgc acacagcagg
     4321 catatcagcc tcagtagccc ttctaatatc agcaggcctt ctttcaggag acatgcgaaa
     4381 cgatccagat ccggtgcaga ttatttggat tgagagtgaa tatgagactc taattggata
     4441 ccgaggggaa tttatggaac gtcagtggag cattttgaca agaaatattt gctagctgat
     4501 agtgacctta ggcgactttt gaacgcgcaa taatggtttc tgacgtatgt gcttagctca
     4561 ttaaactcca gaaacccgcg gctgagtggc tccttcaacg ttgcggttct gtcagttcca
     4621 aacgtaaaac ggcttgtccc gcgtcatcgg cgggggtcat aacgtgactc ccttaattct
     4681 ccgctccgga tccgaattcg gcacagagtg ttcaacccca gtacattaaa aacgtccgca
     4741 atgtgttatt aagttgtcta agcgtcaatt tgtttacacc acaatatatc ctgccaccag
     4801 ccagccaaca gctccccgac cggcagctcg gcacaaaatc accactcgat acaggcagcc
     4861 catcaaccat gactactgtc cggtccaaca gatccgagca ggaaccgtaa aaaggccgcg
     4921 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4981 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5041 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5101 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5161 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5221 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5281 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5341 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5401 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5461 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5521 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5581 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5641 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ctaaaacaat
     5701 tcatccagta aaatataata ttttattttc tcccaatcag gcttgatccc cagtaagtca
     5761 aaaaatagct cgacatactg ttcttccccg atatcctccc tgagcgaccg gacgcagaag
     5821 gcaatgtcat accacttgtc cgccctgccg cttctcccaa gatcaataaa gccacttact
     5881 ttgccatctt tcacaaagat gttgctgtct cccaggtcgc cgtgggaaaa gacaagttcc
     5941 tcttcgggct tttccgtctt taaaaaatca tacagctcgc gcggatcttt aaatggagtg
     6001 tcctcttccc agttttcgca atccacatcg gccagatcgt tattcagtaa gtaatccaat
     6061 tcggctaagc ggctgtctaa gctattcgta tagggacaat ccgatatgtc gatggagtga
     6121 aagagcctga tgcactccgc atacagctcg ataatctttt cagggctttg ttcatcttca
     6181 tactcttccg agcaaaggac gccatcggcc tcactcatga gcagattgct ccagccatca
     6241 tgccgttcaa agtgcaggac ctttggaaca ggcagctttc cttccagcca tagcatcatg
     6301 tccttttccc gttccacatc ataggtggtc cctttatacc ggctgtccgt catttttaaa
     6361 tataggtttt cattttctcc caccagctta tataccttag caggagacat tccttccgta
     6421 tcttttacgc agcggtattt ttcgatcagt tttttcaatt ccggtgatat tctcatttta
     6481 gccatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     6541 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cactcggagc
     6601 cttgatgtgg gcgctttttg cagctgttcg gctgtgcgct ggccagacag ttatgcacag
     6661 gccaggcggg ttttaagagt tttaataagt tttaaagagt tttaggcgga aaaatcgcct
     6721 tttttctctt ttatatcagt cacttacatg tgtgaccggt tcccaatgta cggctttggg
     6781 ttcccaatgt acgggttccg gttcccaatg tacggctttg ggttcccaat gtacgtgcta
     6841 tccacaggaa agaagctttt tcgacctttt tcccctgcta gggcaatttg ccctagcatc
     6901 tgctccgtac attaggaacc ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg
     6961 actaggatcg ggccagcctg ccccgcctcc tccttcaaat cgtactccgg caggtcattt
     7021 gacccgatca gcttgcgcac ggtgaaacag aacttcttga actctccggc gctgccactg
     7081 cgttcgtaga tcgtcttgaa caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc
     7141 aggcggtaga gaaaacggcc gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc
     7201 agcacgtccg ggttcttgcc ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc
     7261 tcgatgtact ccggccgccc ggtttcgctc tttacgatct tgtagcggct aatcaaggct
     7321 tcaccctcgg ataccgtcac caggcggccg ttcttggcct tcttcgtacg ctgcatggca
     7381 acgtgcgtgg tgtttaaccg aatgcaggtt tctaccaggt cgtctttctg ctttccgcca
     7441 tcggctcgcc ggcagaactt gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc
     7501 ttgtctccct tcccttcccg gtatcggttc atggattcgg ttagatggga aaccgccatc
     7561 agtaccaggt cgtaatccca cacactggcc atgccggccg gccctgcgga aacctctacg
     7621 tgcccgtctg gaagctcgta gcggatcacc tcgccagctc gtcggtcacg cttcgacaga
     7681 cggaaaacgg ccacgtccat gatgctgcga ctatcgcggg tgcccacgtc atagagcatc
     7741 ggaacgaaaa aatctggttg ctcgtcgccc ttgggcggct tcctaatcga cgacgcaccg
     7801 gctgccggcg gttgccggga ttctttgcgg attcgatcag cggccgcttg ccacgattca
     7861 ccggggcgtg cttctgcctc gatgcgttgc cgctgggcgg cctgcgcggc cttcaacttc
     7921 tccaccaggt catcacccag cgccgcgccg atttgtaccg ggccggatgg tttgcgaccg
     7981 ctcacgccga ttcctcgggc ttgggggttc cagtgccatt gcagggccgg cagacaaccc
     8041 agccgcttac gcctggccaa ccgcccgttc ctccacacat ggggcattcc acggcgtcgg
     8101 tgcctggttg ttcttgattt tccatgccgc ctcctttagc cgctaaaatt catctactca
     8161 tttattcatt tgctcattta ctctggtagc tgcgcgatgt attcagatag cagctcggta
     8221 atggtcttgc cttggcgtac cg
//
