LOCUS       sequence-342112-        8302 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 8302)
  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 8302)
  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_02860a
FEATURES             Location/Qualifiers
     source          1..8302
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    635..659
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(726..1433)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     promoter        complement(1750..2095)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(1787..1807)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      complement(3128..3380)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        complement(3980..4162)
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        complement(4016..4194)
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    4357..4381
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4517..5105)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4597..4616)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5276..6070)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     promoter        complement(6071..6175)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6237..6431
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(6497..7570)
                     /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(7999..8302,1..326))
                     /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 gggcttggct gcaccggaat cagcacgaag tcggctgcct tgatcgcgga cacagccaag
       61 tccgccgcct ggggcgctcc gtcgatcact acgaagtcgc gccggccgat ggccttcacg
      121 tcgcggtcaa tcgtcgggcg gtcgatgccg acaacggtta gcggttgatc ttcccgcacg
      181 gccgcccaat cgcgggcact gccctgggga tcggaatcga ctaacagaac atcggccccg
      241 gcgagttgca gggcgcgggc tagatgggtt gcgatggtcg tcttgcctga cccgcctttc
      301 tggttaagta cagcgataac cttcatgcgt tccccttgcg tatttgttta tttactcatc
      361 gcatcatata cgcagcgacc gcatgacgca agctgtttta ctcaaataca catcaccttt
      421 ttagacggcg gcgctcggtt tcttcagcgg ccaagctggc cggccaggcc gccagcttgg
      481 catcagacaa accggccagg atttcatgca gccgcacggc tgactcccga ggggaaccct
      541 gtggttggca tgcacataca aatggacgaa cggataaacc ttttcacgcc cttttaaata
      601 tccgattatt ctaataaacg ctcttttctc ttaggtttac ccgccaatat atcctgtcaa
      661 acactgatag tttaaacgga tcacttcgtg tccctcggtc acatgtgcat cctctagtag
      721 cggtcctgct gagcctcgac atgttgtcgc aaaattcgcc ctggacccgc ccaacgattt
      781 gtcgtcactg tcaaggtttg acctgcactt catttggggc ccacatacac caaaaaaatg
      841 ctgcataatt ctcggggcag caagtcggtt acccggccgc cgtgctggac cgggttgaat
      901 ggtgcccgta actttcggta gagcggacgg ccaatactca acttcaagga atctcaccca
      961 tgcgcgccgg cggggaaccg gagttccctt cagtgagcgt tattagttcg ccgctcggtg
     1021 tgtcgtagat actagcccct ggggcacttt tgaaatttga ataagattta tgtaatcagt
     1081 cttttaggtt tgaccggttc tgccgctttt tttaaaattg gatttgtaat aataaaacgc
     1141 aattgtttgt tattgtggcg ctctatcata gatgtcgcta taaacctatt cagcacaata
     1201 tattgttttc attttaatat tgtacatata agtagtaggg tacaatcagt aaattgaacg
     1261 gagaatatta ttcataaaaa tacgatagta acgggtgata tattcattag aatgaaccga
     1321 aaccggcggt aaggatctga gctacacatg ctcaggtttt ttacaacgtg cacaacagaa
     1381 ttgaaagcaa atatcatgcg atcataggct tctcgcatat ctcattaaag caggacaagc
     1441 tcaaggtccc ctgaaagcct gagggtcctt caagtactta gtgtacaagt tagagatcct
     1501 agcagcagtg tcgatagggt atccagacct gaagatctcg tcgtacacgt caagagcctg
     1561 agacttcttc tgcaagaaag cagagaacat tgatgaaagc tccttgatag cagccacctc
     1621 gtcgtcgtcc aacctgaaca actcagacaa cttagtcatt gtatcgataa ttgtaaatgt
     1681 aattgtaatg ttgtttgttg tttgttgttg ttggtaattg ttgtaaaaat gagctcttat
     1741 actcgagcgt gtcctctcca aatgaaatga acttccttat atagaggaag ggtcttgcga
     1801 aggatagtgg gattgtgcgt catcccttac gtcagtggag atgtcacatc aatccacttg
     1861 ctttgtagac gtggttggaa cctcttcttt ttccacgatg ctcctcgtgg gtgggggtcc
     1921 atctttggga ccactgtcgg cagagagatc ttgaatgata gcctttcctt tatcgcaatg
     1981 atggcatttg taggagccac cttccttttc tactgtcctt tcgatgaagt gacagatagc
     2041 tgggcaatgg aatccgagga ggtttcccga aattatcctt tgttgaaaag tctcaatagc
     2101 cctttgatct tctgagactg tatctttgac atttttggag tagaccagag tgtcgtgctc
     2161 caccatgttg acgaagattt tcttcttgtc attgagtcgt aaaagactct gtatgaactg
     2221 ttcgccagtc ttgacggcga gttctgttag atcctcgatt tgaatcttag actccatgca
     2281 tggccttaga ttcagtagga actacctttt tagagactcc aatctctatt acttgccttg
     2341 gtttatgaag caagccttga atcgtccata ctggaatagt acttctgatc ttgagaaata
     2401 tgtctttctc tgtgttcttg atgcaattag tcctgaatct tttgactgca tctttaacct
     2461 tcttgggaag gtatttgatc tcctggagat tgttactcgg gtagatcgtc ttgatgagac
     2521 ttgctgcgta ggcctctcta accatctgtg ggtcagcatt ctttctgaaa ttgaagaggc
     2581 taaccttctc attatcagtg gtgaacatag tgtcgtcacc ttcaccttcg aacttccttc
     2641 ctagatcgta aagatagagg aaatcgtcca ttgtaatctc cggggcaaag gagatctctt
     2701 ttggggctgg atcactgctg ggccttttgg ttcctagcgt gagccagtgg gctttttgct
     2761 ttggtgggct tgttagggcc ttagcaaagc tcttgggctt gagttgagct tctcctttgg
     2821 ggatgaagtt caacctgtct gtttgctgac ttgttgtgta cgcgtcagct gctgctcttg
     2881 cctctgtaat agtggcaaat ttcttgtgtg caactccggg aacgccgttt gttgccgcct
     2941 ttgtacaacc ccagtcatcg tatataccgg catgtggacc gttatacaca acgtagtagt
     3001 tgatatgagg gtgttgaata cccgattctg ctctgagagg agcaactgtg ctgttaagct
     3061 cagatttttg tgggattgga attcctccaa gcttgaattc ttgcagcgct ccaattcagc
     3121 aagcgtcgat ctagtaacat agatgacacc gcgcgcgata atttatccta gtttgcgcgc
     3181 tatattttgt tttctatcgc gtattaaatg tataattgcg ggactctaat cataaaaacc
     3241 catctcataa ataacgtcat gcattacatg ttaattatta catgcttaac gtaattcaac
     3301 agaaattata tgataatcat cgcaagaccg gcaacaggat tcaatcttaa gaaactttat
     3361 tgccaaatgt ttgaacgatc tgcttgactc tagatccaga gtcccgctca aatctcagtc
     3421 acaggaagaa caggtctagg aggaacagga agagagaaat caagctgcca gaatcccaca
     3481 tcatgccaat ttccatgttt gaaaccagca gctctaagca ttcccctagg agcataacca
     3541 agagcctcat gcatcctcac agaaggatca ttaggaagtc caatcacagc aaccacagac
     3601 ttgaaaccct gagcctccaa agacttcaaa agatgagtgt aaagagtaga tccaagacca
     3661 gttctttgat gcctaggaga cacgtaaaca gtagactcag cagtccaatc gtaagcattc
     3721 ctagccttcc aaggaccagc ataagcaata ccagccactt caccatccac ttcagcaaca
     3781 agccaaggat acctctctct aagcctcaca agatcatcag tccactcttg tggttcttga
     3841 ggctcagtcc tgaaattcac agtagaagtc tcaatgtaat gattcacaat agtgcacaca
     3901 gcaggcatat cagcctcagt agcccttcta atatcagcag gccttctttc aggagacatg
     3961 cgaaacgatc cagatccggt gcagattatt tggattgaga gtgaatatga gactctaatt
     4021 ggataccgag gggaatttat ggaacgtcag tggagcattt tgacaagaaa tatttgctag
     4081 ctgatagtga ccttaggcga cttttgaacg cgcaataatg gtttctgacg tatgtgctta
     4141 gctcattaaa ctccagaaac ccgcggctga gtggctcctt caacgttgcg gttctgtcag
     4201 ttccaaacgt aaaacggctt gtcccgcgtc atcggcgggg gtcataacgt gactccctta
     4261 attctccgct ccggatccga attcggcaca gagtgttcaa ccccagtaca ttaaaaacgt
     4321 ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt acaccacaat atatcctgcc
     4381 accagccagc caacagctcc ccgaccggca gctcggcaca aaatcaccac tcgatacagg
     4441 cagcccatca accatgacta ctgtccggtc caacagatcc gagcaggaac cgtaaaaagg
     4501 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4561 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4621 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4681 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4741 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4801 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4861 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4921 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4981 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     5041 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5101 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5161 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5221 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagctaaa
     5281 acaattcatc cagtaaaata taatatttta ttttctccca atcaggcttg atccccagta
     5341 agtcaaaaaa tagctcgaca tactgttctt ccccgatatc ctccctgagc gaccggacgc
     5401 agaaggcaat gtcataccac ttgtccgccc tgccgcttct cccaagatca ataaagccac
     5461 ttactttgcc atctttcaca aagatgttgc tgtctcccag gtcgccgtgg gaaaagacaa
     5521 gttcctcttc gggcttttcc gtctttaaaa aatcatacag ctcgcgcgga tctttaaatg
     5581 gagtgtcctc ttcccagttt tcgcaatcca catcggccag atcgttattc agtaagtaat
     5641 ccaattcggc taagcggctg tctaagctat tcgtataggg acaatccgat atgtcgatgg
     5701 agtgaaagag cctgatgcac tccgcataca gctcgataat cttttcaggg ctttgttcat
     5761 cttcatactc ttccgagcaa aggacgccat cggcctcact catgagcaga ttgctccagc
     5821 catcatgccg ttcaaagtgc aggacctttg gaacaggcag ctttccttcc agccatagca
     5881 tcatgtcctt ttcccgttcc acatcatagg tggtcccttt ataccggctg tccgtcattt
     5941 ttaaatatag gttttcattt tctcccacca gcttatatac cttagcagga gacattcctt
     6001 ccgtatcttt tacgcagcgg tatttttcga tcagtttttt caattccggt gatattctca
     6061 ttttagccat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6121 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcactc
     6181 ggagccttga tgtgggcgct ttttgcagct gttcggctgt gcgctggcca gacagttatg
     6241 cacaggccag gcgggtttta agagttttaa taagttttaa agagttttag gcggaaaaat
     6301 cgcctttttt ctcttttata tcagtcactt acatgtgtga ccggttccca atgtacggct
     6361 ttgggttccc aatgtacggg ttccggttcc caatgtacgg ctttgggttc ccaatgtacg
     6421 tgctatccac aggaaagaag ctttttcgac ctttttcccc tgctagggca atttgcccta
     6481 gcatctgctc cgtacattag gaaccggcgg atgcttcgcc ctcgatcagg ttgcggtagc
     6541 gcatgactag gatcgggcca gcctgccccg cctcctcctt caaatcgtac tccggcaggt
     6601 catttgaccc gatcagcttg cgcacggtga aacagaactt cttgaactct ccggcgctgc
     6661 cactgcgttc gtagatcgtc ttgaacaacc atctggcttc tgccttgcct gcggcgcggc
     6721 gtgccaggcg gtagagaaaa cggccgatgc cgggatcgat caaaaagtaa tcggggtgaa
     6781 ccgtcagcac gtccgggttc ttgccttctg tgatctcgcg gtacatccaa tcagctagct
     6841 cgatctcgat gtactccggc cgcccggttt cgctctttac gatcttgtag cggctaatca
     6901 aggcttcacc ctcggatacc gtcaccaggc ggccgttctt ggccttcttc gtacgctgca
     6961 tggcaacgtg cgtggtgttt aaccgaatgc aggtttctac caggtcgtct ttctgctttc
     7021 cgccatcggc tcgccggcag aacttgagta cgtccgcaac gtgtggacgg aacacgcggc
     7081 cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga tgggaaaccg
     7141 ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct gcggaaacct
     7201 ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg tcacgcttcg
     7261 acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc acgtcataga
     7321 gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta atcgacgacg
     7381 caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc gcttgccacg
     7441 attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc gcggccttca
     7501 acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg gatggtttgc
     7561 gaccgctcac gccgattcct cgggcttggg ggttccagtg ccattgcagg gccggcagac
     7621 aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatggggc attccacggc
     7681 gtcggtgcct ggttgttctt gattttccat gccgcctcct ttagccgcta aaattcatct
     7741 actcatttat tcatttgctc atttactctg gtagctgcgc gatgtattca gatagcagct
     7801 cggtaatggt cttgccttgg cgtaccgcgt acatcttcag cttggtgtga tcctccgccg
     7861 gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc caggctggcc aacgttgcag
     7921 ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt gtttgtgctt ttgctcattt
     7981 tctctttacc tcattaactc aaatgagttt tgatttaatt tcagcggcca gcgcctggac
     8041 ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg gttgtgccgg cggcggcagt
     8101 gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga atgggcagct cgtacccggc
     8161 cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg atcgcccgcg acacgacaaa
     8221 ggccgcttgt agccttccat ccgtgacctc aatgcgctgc ttaaccagct ccaccaggtc
     8281 ggcggtggcc catatgtcgt aa
//
