LOCUS       sequence-342546-        9469 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 9469)
  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 9469)
  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_15278a
FEATURES             Location/Qualifiers
     source          1..9469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1198..1543
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1486..1506
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      3027..3734
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    3801..3825
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4134..4763
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5192..6265
                     /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"
     rep_origin      6331..6525
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     promoter        6587..6691
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6692..7486
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      7657..8245
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8146..8165
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    8381..8405
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        8568..8746
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        8600..8782
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     terminator      join(9382..9469,1..165)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 acgttaagca tgtaataatt aacatgtaat gcatgacgtt atttatgaga tgggttttta
       61 tgattagagt cccgcaatta tacatttaat acgcgataga aaacaaaata tagcgcgcaa
      121 actaggataa attatcgcgc gcggtgtcat ctatgttact agatcgacgc ttgctgaatt
      181 ggagcgctgc aagaattcaa gcttggagga attccaatcc cacaaaaatc tgagcttaac
      241 agcacagttg ctcctctcag agcagaatcg ggtattcaac accctcatat caactactac
      301 gttgtgtata acggtccaca tgccggtata tacgatgact ggggttgtac aaaggcggca
      361 acaaacggcg ttcccggagt tgcacacaag aaatttgcca ctattacaga ggcaagagca
      421 gcagctgacg cgtacacaac aagtcagcaa acagacaggt tgaacttcat ccccaaagga
      481 gaagctcaac tcaagcccaa gagctttgct aaggccctaa caagcccacc aaagcaaaaa
      541 gcccactggc tcacgctagg aaccaaaagg cccagcagtg atccagcccc aaaagagatc
      601 tcctttgccc cggagattac aatggacgat ttcctctatc tttacgatct aggaaggaag
      661 ttcgaaggtg aaggtgacga cactatgttc accactgata atgagaaggt tagcctcttc
      721 aatttcagaa agaatgctga cccacagatg gttagagagg cctacgcagc aagtctcatc
      781 aagacgatct acccgagtaa caatctccag gagatcaaat accttcccaa gaaggttaaa
      841 gatgcagtca aaagattcag gactaattgc atcaagaaca cagagaaaga catatttctc
      901 aagatcagaa gtactattcc agtatggacg attcaaggct tgcttcataa accaaggcaa
      961 gtaatagaga ttggagtctc taaaaaggta gttcctactg aatctaaggc catgcatgga
     1021 gtctaagatt caaatcgagg atctaacaga actcgccgtc aagactggcg aacagttcat
     1081 acagagtctt ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga
     1141 cactctggtc tactccaaaa atgtcaaaga tacagtctca gaagatcaaa gggctattga
     1201 gacttttcaa caaaggataa tttcgggaaa cctcctcgga ttccattgcc cagctatctg
     1261 tcacttcatc gaaaggacag tagaaaagga aggtggctcc tacaaatgcc atcattgcga
     1321 taaaggaaag gctatcattc aagatctctc tgccgacagt ggtcccaaag atggaccccc
     1381 acccacgagg agcatcgtgg aaaaagaaga ggttccaacc acgtctacaa agcaagtgga
     1441 ttgatgtgac atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga
     1501 cccttcctct atataaggaa gttcatttca tttggagagg acacgctcga gtataagagc
     1561 tcatttttac aacaattacc aacaacaaca aacaacaaac aacattacaa ttacatttac
     1621 aattatcgat acaatgggat tcccatcagg accatcactt gaaaagattg aagctatgtt
     1681 tcagtcattg actaacaaga tcacaacaaa gtctaagcca tcacagaggg acgtgccaga
     1741 cgaaaagttg tctcagcttt tgggtgctgc aaagcctgtg aacaaggcag ttgaggcagc
     1801 tggtaggctt aagactttgc aaactcagaa gtggttggac gagggaaagt caactcagga
     1861 agtgttccag ttgttggaat tgaacaagtt catggtgcct aacttcaaga agatcgaggg
     1921 aactgtgttt ggattgcctg acttcaatac atgggttaat tacgttgacg actttaacgc
     1981 aaaaaaccca actaagaaag agtctatgat cccaacattg aggactatct attctgacga
     2041 gggtttgaca agagctcttg agttggctaa gacatactct actacaaacg cattggctac
     2101 taagttgagg aaagagcaga tccaaagatg gttggacgac gcacagacac caaaatatgt
     2161 gtttgagatg ttcatgatcg acgacaaggt ggacggattg ttgacaaacc ctaggtttat
     2221 cgcttggaca aagtacgtgg atgacttcaa ccttcagtac ccaacatcta gggcttcaat
     2281 ggagcctcca attgctgctc actacggtga cgacgcagtg ttcgctatgt tggaggcagc
     2341 taaaaaggtg cagcttacag agaacgtggc ttcaaggttg caggctgagc agatcaaaag
     2401 gcttcttaac tctaacagat ctccagagta tgtgtttgaa gctttcaatc ttcatgagac
     2461 tggtgataac cttttgtcaa caccaatgtt caagacatgg ttcaattatt tggagtcatt
     2521 taacaagaaa aacactgata aggagacttt gttggctcct atccacaggt actaccacga
     2581 ccacggagtg gctaagatcg tggctgaggg tatgaagaat ccatctactg ttgaattgtc
     2641 aaaacagctt cagatccaga gatacaagag gtggttgcat gctgagaggc cacctaggga
     2701 cgcattcaac acattcatcc ttgaaaaacc aggagctgat gtgattatca ggtacaacga
     2761 gaggttggac ggaaagttgt accaagttag attgaataag gtgtcagacg gacttctttc
     2821 atctccagag tttcagcttt ggtctaagta tttggatgat tggaatacaa agtacccaga
     2881 ccagaaacaa tctatggcta aaatcttcca ggcacttttc actgaggacg cacttgctaa
     2941 gatgatcatg gctgcaagga acaacccatc aactcagaaa atcgcttctt tgcttgaaaa
     3001 cgcatttgca aaggtttgag cttgtcctgc tttaatgaga tatgcgagaa gcctatgatc
     3061 gcatgatatt tgctttcaat tctgttgtgc acgttgtaaa aaacctgagc atgtgtagct
     3121 cagatcctta ccgccggttt cggttcattc taatgaatat atcacccgtt actatcgtat
     3181 ttttatgaat aatattctcc gttcaattta ctgattgtac cctactactt atatgtacaa
     3241 tattaaaatg aaaacaatat attgtgctga ataggtttat agcgacatct atgatagagc
     3301 gccacaataa caaacaattg cgttttatta ttacaaatcc aattttaaaa aaagcggcag
     3361 aaccggtcaa acctaaaaga ctgattacat aaatcttatt caaatttcaa aagtgcccca
     3421 ggggctagta tctacgacac accgagcggc gaactaataa cgctcactga agggaactcc
     3481 ggttccccgc cggcgcgcat gggtgagatt ccttgaagtt gagtattggc cgtccgctct
     3541 accgaaagtt acgggcacca ttcaacccgg tccagcacgg cggccgggta accgacttgc
     3601 tgccccgaga attatgcagc atttttttgg tgtatgtggg ccccaaatga agtgcaggtc
     3661 aaaccttgac agtgacgaca aatcgttggg cgggtccagg gcgaattttg cgacaacatg
     3721 tcgaggctca gcaggaccgc tactagagga tgcacatgtg accgagggac acgaagtgat
     3781 ccgtttaaac tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc
     3841 gtttattaga ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt
     3901 gtatgtgcat gccaaccaca gggttcccct cgggagtcag ccgtgcggct gcatgaaatc
     3961 ctggccggtt tgtctgatgc caagctggcg gcctggccgg ccagcttggc cgctgaagaa
     4021 accgagcgcc gccgtctaaa aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg
     4081 gtcgctgcgt atatgatgcg atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg
     4141 ttatcgctgt acttaaccag aaaggcgggt caggcaagac gaccatcgca acccatctag
     4201 cccgcgccct gcaactcgcc ggggccgatg ttctgttagt cgattccgat ccccagggca
     4261 gtgcccgcga ttgggcggcc gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc
     4321 gcccgacgat tgaccgcgac gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg
     4381 gagcgcccca ggcggcggac ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga
     4441 ttccggtgca gccaagccct tacgacatat gggccaccgc cgacctggtg gagctggtta
     4501 agcagcgcat tgaggtcacg gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga
     4561 tcaaaggcac gcgcatcggc ggtgaggttg ccgaggcgct ggccgggtac gagctgccca
     4621 ttcttgagtc ccgtatcacg cagcgcgtga gctacccagg cactgccgcc gccggcacaa
     4681 ccgttcttga atcagaaccc gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg
     4741 aaattaaatc aaaactcatt tgagttaatg aggtaaagag aaaatgagca aaagcacaaa
     4801 cacgctaagt gccggccgtc cgagcgcacg cagcagcaag gctgcaacgt tggccagcct
     4861 ggcagacacg ccagccatga agcgggtcaa ctttcagttg ccggcggagg atcacaccaa
     4921 gctgaagatg tacgcggtac gccaaggcaa gaccattacc gagctgctat ctgaatacat
     4981 cgcgcagcta ccagagtaaa tgagcaaatg aataaatgag tagatgaatt ttagcggcta
     5041 aaggaggcgg catggaaaat caagaacaac caggcaccga cgccgtggaa tgccccatgt
     5101 gtggaggaac gggcggttgg ccaggcgtaa gcggctgggt tgtctgccgg ccctgcaatg
     5161 gcactggaac ccccaagccc gaggaatcgg cgtgagcggt cgcaaaccat ccggcccggt
     5221 acaaatcggc gcggcgctgg gtgatgacct ggtggagaag ttgaaggccg cgcaggccgc
     5281 ccagcggcaa cgcatcgagg cagaagcacg ccccggtgaa tcgtggcaag cggccgctga
     5341 tcgaatccgc aaagaatccc ggcaaccgcc ggcagccggt gcgtcgtcga ttaggaagcc
     5401 gcccaagggc gacgagcaac cagatttttt cgttccgatg ctctatgacg tgggcacccg
     5461 cgatagtcgc agcatcatgg acgtggccgt tttccgtctg tcgaagcgtg accgacgagc
     5521 tggcgaggtg atccgctacg agcttccaga cgggcacgta gaggtttccg cagggccggc
     5581 cggcatggcc agtgtgtggg attacgacct ggtactgatg gcggtttccc atctaaccga
     5641 atccatgaac cgataccggg aagggaaggg agacaagccc ggccgcgtgt tccgtccaca
     5701 cgttgcggac gtactcaagt tctgccggcg agccgatggc ggaaagcaga aagacgacct
     5761 ggtagaaacc tgcattcggt taaacaccac gcacgttgcc atgcagcgta cgaagaaggc
     5821 caagaacggc cgcctggtga cggtatccga gggtgaagcc ttgattagcc gctacaagat
     5881 cgtaaagagc gaaaccgggc ggccggagta catcgagatc gagctagctg attggatgta
     5941 ccgcgagatc acagaaggca agaacccgga cgtgctgacg gttcaccccg attacttttt
     6001 gatcgatccc ggcatcggcc gttttctcta ccgcctggca cgccgcgccg caggcaaggc
     6061 agaagccaga tggttgttca agacgatcta cgaacgcagt ggcagcgccg gagagttcaa
     6121 gaagttctgt ttcaccgtgc gcaagctgat cgggtcaaat gacctgccgg agtacgattt
     6181 gaaggaggag gcggggcagg ctggcccgat cctagtcatg cgctaccgca acctgatcga
     6241 gggcgaagca tccgccggtt cctaatgtac ggagcagatg ctagggcaaa ttgccctagc
     6301 aggggaaaaa ggtcgaaaaa gcttctttcc tgtggatagc acgtacattg ggaacccaaa
     6361 gccgtacatt gggaaccgga acccgtacat tgggaaccca aagccgtaca ttgggaaccg
     6421 gtcacacatg taagtgactg atataaaaga gaaaaaaggc gatttttccg cctaaaactc
     6481 tttaaaactt attaaaactc ttaaaacccg cctggcctgt gcataactgt ctggccagcg
     6541 cacagccgaa cagctgcaaa aagcgcccac atcaaggctc cgagtgcgcg gaacccctat
     6601 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6661 aatgcttcaa taatattgaa aaaggaagag tatggctaaa atgagaatat caccggaatt
     6721 gaaaaaactg atcgaaaaat accgctgcgt aaaagatacg gaaggaatgt ctcctgctaa
     6781 ggtatataag ctggtgggag aaaatgaaaa cctatattta aaaatgacgg acagccggta
     6841 taaagggacc acctatgatg tggaacggga aaaggacatg atgctatggc tggaaggaaa
     6901 gctgcctgtt ccaaaggtcc tgcactttga acggcatgat ggctggagca atctgctcat
     6961 gagtgaggcc gatggcgtcc tttgctcgga agagtatgaa gatgaacaaa gccctgaaaa
     7021 gattatcgag ctgtatgcgg agtgcatcag gctctttcac tccatcgaca tatcggattg
     7081 tccctatacg aatagcttag acagccgctt agccgaattg gattacttac tgaataacga
     7141 tctggccgat gtggattgcg aaaactggga agaggacact ccatttaaag atccgcgcga
     7201 gctgtatgat tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt cccacggcga
     7261 cctgggagac agcaacatct ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg
     7321 gagaagcggc agggcggaca agtggtatga cattgccttc tgcgtccggt cgctcaggga
     7381 ggatatcggg gaagaacagt atgtcgagct attttttgac ttactgggga tcaagcctga
     7441 ttgggagaaa ataaaatatt atattttact ggatgaattg ttttagctgt cagaccaagt
     7501 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     7561 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     7621 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     7681 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     7741 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     7801 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     7861 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     7921 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     7981 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     8041 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     8101 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     8161 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     8221 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctgct
     8281 cggatctgtt ggaccggaca gtagtcatgg ttgatgggct gcctgtatcg agtggtgatt
     8341 ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg
     8401 taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg
     8461 gttgaacact ctgtgccgaa ttcggatccg gagcggagaa ttaagggagt cacgttatga
     8521 cccccgccga tgacgcggga caagccgttt tacgtttgga actgacagaa ccgcaacgtt
     8581 gaaggagcca ctcagccgcg ggtttctgga gtttaatgag ctaagcacat acgtcagaaa
     8641 ccattattgc gcgttcaaaa gtcgcctaag gtcactatca gctagcaaat atttcttgtc
     8701 aaaatgctcc actgacgttc cataaattcc cctcggtatc caattagagt ctcatattca
     8761 ctctcaatcc aaataatctg caccggatct ggatcgtttc gcatgtctcc tgaaagaagg
     8821 cctgctgata ttagaagggc tactgaggct gatatgcctg ctgtgtgcac tattgtgaat
     8881 cattacattg agacttctac tgtgaatttc aggactgagc ctcaagaacc acaagagtgg
     8941 actgatgatc ttgtgaggct tagagagagg tatccttggc ttgttgctga agtggatggt
     9001 gaagtggctg gtattgctta tgctggtcct tggaaggcta ggaatgctta cgattggact
     9061 gctgagtcta ctgtttacgt gtctcctagg catcaaagaa ctggtcttgg atctactctt
     9121 tacactcatc ttttgaagtc tttggaggct cagggtttca agtctgtggt tgctgtgatt
     9181 ggacttccta atgatccttc tgtgaggatg catgaggctc ttggttatgc tcctagggga
     9241 atgcttagag ctgctggttt caaacatgga aattggcatg atgtgggatt ctggcagctt
     9301 gatttctctc ttcctgttcc tcctagacct gttcttcctg tgactgagat ttgagcggga
     9361 ctctggatct agagtcaagc agatcgttca aacatttggc aataaagttt cttaagattg
     9421 aatcctgttg ccggtcttgc gatgattatc atataatttc tgttgaatt
//
