LOCUS       sequence-342754-        8341 bp    DNA     circular SYN 15-APR-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 8341)
  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 8341)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_17063
FEATURES             Location/Qualifiers
     source          1..8341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    753..777
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(844..1551)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1907..2252)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1944..1964)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3285..3537)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(4137..4319)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4173..4351)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4514..4538
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4674..5262)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4754..4773)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5433..6227)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6228..6332)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6394..6588
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6654..7727)
                     /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(8156..8341,1..444))
                     /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 gatgcgcgtg cctttgatcg cccgcgacac gacaaaggcc gcttgtagcc ttccatccgt
       61 gacctcaatg cgctgcttaa ccagctccac caggtcggcg gtggcccata tgtcgtaagg
      121 gcttggctgc accggaatca gcacgaagtc ggctgccttg atcgcggaca cagccaagtc
      181 cgccgcctgg ggcgctccgt cgatcactac gaagtcgcgc cggccgatgg ccttcacgtc
      241 gcggtcaatc gtcgggcggt cgatgccgac aacggttagc ggttgatctt cccgcacggc
      301 cgcccaatcg cgggcactgc cctggggatc ggaatcgact aacagaacat cggccccggc
      361 gagttgcagg gcgcgggcta gatgggttgc gatggtcgtc ttgcctgacc cgcctttctg
      421 gttaagtaca gcgataacct tcatgcgttc cccttgcgta tttgtttatt tactcatcgc
      481 atcatatacg cagcgaccgc atgacgcaag ctgttttact caaatacaca tcaccttttt
      541 agacggcggc gctcggtttc ttcagcggcc aagctggccg gccaggccgc cagcttggca
      601 tcagacaaac cggccaggat ttcatgcagc cgcacggctg actcccgagg ggaaccctgt
      661 ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct tttaaatatc
      721 cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat cctgtcaaac
      781 actgatagtt taaacggatc acttcgtgtc cctcggtcac atgtgcatcc tctagtagcg
      841 gtcctgctga gcctcgacat gttgtcgcaa aattcgccct ggacccgccc aacgatttgt
      901 cgtcactgtc aaggtttgac ctgcacttca tttggggccc acatacacca aaaaaatgct
      961 gcataattct cggggcagca agtcggttac ccggccgccg tgctggaccg ggttgaatgg
     1021 tgcccgtaac tttcggtaga gcggacggcc aatactcaac ttcaaggaat ctcacccatg
     1081 cgcgccggcg gggaaccgga gttcccttca gtgagcgtta ttagttcgcc gctcggtgtg
     1141 tcgtagatac tagcccctgg ggcacttttg aaatttgaat aagatttatg taatcagtct
     1201 tttaggtttg accggttctg ccgctttttt taaaattgga tttgtaataa taaaacgcaa
     1261 ttgtttgtta ttgtggcgct ctatcataga tgtcgctata aacctattca gcacaatata
     1321 ttgttttcat tttaatattg tacatataag tagtagggta caatcagtaa attgaacgga
     1381 gaatattatt cataaaaata cgatagtaac gggtgatata ttcattagaa tgaaccgaaa
     1441 ccggcggtaa ggatctgagc tacacatgct caggtttttt acaacgtgca caacagaatt
     1501 gaaagcaaat atcatgcgat cataggcttc tcgcatatct cattaaagca ggacaagctc
     1561 acttagccaa gtgagatctt ccagtcctgt gcaaccactc agagtacaac ctagcgtacc
     1621 tcttaaatcc agaaggagtt ccccaccttc cagagttcat catcctcacc ctgaaagtgt
     1681 ccctcaactc agagtaagtc actcccttct cagccatctt cttgaatcct ggcaccaaca
     1741 ccttctgctc ccacttcaag tacctagcct tgatagcctg aagagcctta gtcaacttct
     1801 gagacactcc cttcattgta tcgataattg taaatgtaat tgtaatgttg tttgttgttt
     1861 gttgttgttg gtaattgttg taaaaatgag ctcttatact cgagcgtgtc ctctccaaat
     1921 gaaatgaact tccttatata gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat
     1981 cccttacgtc agtggagatg tcacatcaat ccacttgctt tgtagacgtg gttggaacct
     2041 cttctttttc cacgatgctc ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag
     2101 agagatcttg aatgatagcc tttcctttat cgcaatgatg gcatttgtag gagccacctt
     2161 ccttttctac tgtcctttcg atgaagtgac agatagctgg gcaatggaat ccgaggaggt
     2221 ttcccgaaat tatcctttgt tgaaaagtct caatagccct ttgatcttct gagactgtat
     2281 ctttgacatt tttggagtag accagagtgt cgtgctccac catgttgacg aagattttct
     2341 tcttgtcatt gagtcgtaaa agactctgta tgaactgttc gccagtcttg acggcgagtt
     2401 ctgttagatc ctcgatttga atcttagact ccatgcatgg ccttagattc agtaggaact
     2461 acctttttag agactccaat ctctattact tgccttggtt tatgaagcaa gccttgaatc
     2521 gtccatactg gaatagtact tctgatcttg agaaatatgt ctttctctgt gttcttgatg
     2581 caattagtcc tgaatctttt gactgcatct ttaaccttct tgggaaggta tttgatctcc
     2641 tggagattgt tactcgggta gatcgtcttg atgagacttg ctgcgtaggc ctctctaacc
     2701 atctgtgggt cagcattctt tctgaaattg aagaggctaa ccttctcatt atcagtggtg
     2761 aacatagtgt cgtcaccttc accttcgaac ttccttccta gatcgtaaag atagaggaaa
     2821 tcgtccattg taatctccgg ggcaaaggag atctcttttg gggctggatc actgctgggc
     2881 cttttggttc ctagcgtgag ccagtgggct ttttgctttg gtgggcttgt tagggcctta
     2941 gcaaagctct tgggcttgag ttgagcttct cctttgggga tgaagttcaa cctgtctgtt
     3001 tgctgacttg ttgtgtacgc gtcagctgct gctcttgcct ctgtaatagt ggcaaatttc
     3061 ttgtgtgcaa ctccgggaac gccgtttgtt gccgcctttg tacaacccca gtcatcgtat
     3121 ataccggcat gtggaccgtt atacacaacg tagtagttga tatgagggtg ttgaataccc
     3181 gattctgctc tgagaggagc aactgtgctg ttaagctcag atttttgtgg gattggaatt
     3241 cctccaagct tgaattcttg cagcgctcca attcagcaag cgtcgatcta gtaacataga
     3301 tgacaccgcg cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta
     3361 ttaaatgtat aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca
     3421 ttacatgtta attattacat gcttaacgta attcaacaga aattatatga taatcatcgc
     3481 aagaccggca acaggattca atcttaagaa actttattgc caaatgtttg aacgatctgc
     3541 ttgactctag atccagagtc ccgctcaaat ctcagtcaca ggaagaacag gtctaggagg
     3601 aacaggaaga gagaaatcaa gctgccagaa tcccacatca tgccaatttc catgtttgaa
     3661 accagcagct ctaagcattc ccctaggagc ataaccaaga gcctcatgca tcctcacaga
     3721 aggatcatta ggaagtccaa tcacagcaac cacagacttg aaaccctgag cctccaaaga
     3781 cttcaaaaga tgagtgtaaa gagtagatcc aagaccagtt ctttgatgcc taggagacac
     3841 gtaaacagta gactcagcag tccaatcgta agcattccta gccttccaag gaccagcata
     3901 agcaatacca gccacttcac catccacttc agcaacaagc caaggatacc tctctctaag
     3961 cctcacaaga tcatcagtcc actcttgtgg ttcttgaggc tcagtcctga aattcacagt
     4021 agaagtctca atgtaatgat tcacaatagt gcacacagca ggcatatcag cctcagtagc
     4081 ccttctaata tcagcaggcc ttctttcagg agacatgcga aacgatccag atccggtgca
     4141 gattatttgg attgagagtg aatatgagac tctaattgga taccgagggg aatttatgga
     4201 acgtcagtgg agcattttga caagaaatat ttgctagctg atagtgacct taggcgactt
     4261 ttgaacgcgc aataatggtt tctgacgtat gtgcttagct cattaaactc cagaaacccg
     4321 cggctgagtg gctccttcaa cgttgcggtt ctgtcagttc caaacgtaaa acggcttgtc
     4381 ccgcgtcatc ggcgggggtc ataacgtgac tcccttaatt ctccgctccg gatccgaatt
     4441 cggcacagag tgttcaaccc cagtacatta aaaacgtccg caatgtgtta ttaagttgtc
     4501 taagcgtcaa tttgtttaca ccacaatata tcctgccacc agccagccaa cagctccccg
     4561 accggcagct cggcacaaaa tcaccactcg atacaggcag cccatcaacc atgactactg
     4621 tccggtccaa cagatccgag caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4681 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4741 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4801 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4861 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4921 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4981 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     5041 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     5101 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     5161 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     5221 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     5281 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     5341 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     5401 taaagtatat atgagtaaac ttggtctgac agctaaaaca attcatccag taaaatataa
     5461 tattttattt tctcccaatc aggcttgatc cccagtaagt caaaaaatag ctcgacatac
     5521 tgttcttccc cgatatcctc cctgagcgac cggacgcaga aggcaatgtc ataccacttg
     5581 tccgccctgc cgcttctccc aagatcaata aagccactta ctttgccatc tttcacaaag
     5641 atgttgctgt ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg cttttccgtc
     5701 tttaaaaaat catacagctc gcgcggatct ttaaatggag tgtcctcttc ccagttttcg
     5761 caatccacat cggccagatc gttattcagt aagtaatcca attcggctaa gcggctgtct
     5821 aagctattcg tatagggaca atccgatatg tcgatggagt gaaagagcct gatgcactcc
     5881 gcatacagct cgataatctt ttcagggctt tgttcatctt catactcttc cgagcaaagg
     5941 acgccatcgg cctcactcat gagcagattg ctccagccat catgccgttc aaagtgcagg
     6001 acctttggaa caggcagctt tccttccagc catagcatca tgtccttttc ccgttccaca
     6061 tcataggtgg tccctttata ccggctgtcc gtcattttta aatataggtt ttcattttct
     6121 cccaccagct tatatacctt agcaggagac attccttccg tatcttttac gcagcggtat
     6181 ttttcgatca gttttttcaa ttccggtgat attctcattt tagccatact cttccttttt
     6241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6301 atttagaaaa ataaacaaat aggggttccg cgcactcgga gccttgatgt gggcgctttt
     6361 tgcagctgtt cggctgtgcg ctggccagac agttatgcac aggccaggcg ggttttaaga
     6421 gttttaataa gttttaaaga gttttaggcg gaaaaatcgc cttttttctc ttttatatca
     6481 gtcacttaca tgtgtgaccg gttcccaatg tacggctttg ggttcccaat gtacgggttc
     6541 cggttcccaa tgtacggctt tgggttccca atgtacgtgc tatccacagg aaagaagctt
     6601 tttcgacctt tttcccctgc tagggcaatt tgccctagca tctgctccgt acattaggaa
     6661 ccggcggatg cttcgccctc gatcaggttg cggtagcgca tgactaggat cgggccagcc
     6721 tgccccgcct cctccttcaa atcgtactcc ggcaggtcat ttgacccgat cagcttgcgc
     6781 acggtgaaac agaacttctt gaactctccg gcgctgccac tgcgttcgta gatcgtcttg
     6841 aacaaccatc tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta gagaaaacgg
     6901 ccgatgccgg gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc cgggttcttg
     6961 ccttctgtga tctcgcggta catccaatca gctagctcga tctcgatgta ctccggccgc
     7021 ccggtttcgc tctttacgat cttgtagcgg ctaatcaagg cttcaccctc ggataccgtc
     7081 accaggcggc cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt ggtgtttaac
     7141 cgaatgcagg tttctaccag gtcgtctttc tgctttccgc catcggctcg ccggcagaac
     7201 ttgagtacgt ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc cttcccttcc
     7261 cggtatcggt tcatggattc ggttagatgg gaaaccgcca tcagtaccag gtcgtaatcc
     7321 cacacactgg ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc tggaagctcg
     7381 tagcggatca cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac ggccacgtcc
     7441 atgatgctgc gactatcgcg ggtgcccacg tcatagagca tcggaacgaa aaaatctggt
     7501 tgctcgtcgc ccttgggcgg cttcctaatc gacgacgcac cggctgccgg cggttgccgg
     7561 gattctttgc ggattcgatc agcggccgct tgccacgatt caccggggcg tgcttctgcc
     7621 tcgatgcgtt gccgctgggc ggcctgcgcg gccttcaact tctccaccag gtcatcaccc
     7681 agcgccgcgc cgatttgtac cgggccggat ggtttgcgac cgctcacgcc gattcctcgg
     7741 gcttgggggt tccagtgcca ttgcagggcc ggcagacaac ccagccgctt acgcctggcc
     7801 aaccgcccgt tcctccacac atggggcatt ccacggcgtc ggtgcctggt tgttcttgat
     7861 tttccatgcc gcctccttta gccgctaaaa ttcatctact catttattca tttgctcatt
     7921 tactctggta gctgcgcgat gtattcagat agcagctcgg taatggtctt gccttggcgt
     7981 accgcgtaca tcttcagctt ggtgtgatcc tccgccggca actgaaagtt gacccgcttc
     8041 atggctggcg tgtctgccag gctggccaac gttgcagcct tgctgctgcg tgcgctcgga
     8101 cggccggcac ttagcgtgtt tgtgcttttg ctcattttct ctttacctca ttaactcaaa
     8161 tgagttttga tttaatttca gcggccagcg cctggacctc gcgggcagcg tcgccctcgg
     8221 gttctgattc aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg
     8281 tgatacggga ctcaagaatg ggcagctcgt acccggccag cgcctcggca acctcaccgc
     8341 c
//
