LOCUS       sequence-484647-        5853 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Derivative of pLX-TRV2, a tobacco rattle virus (TRV) vector, 
            designed for the expression of a 32-nt insert to silence CHLI 
            homolog genes in Nicotiana benthamiana and Solanum aethiopicum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5853)
  AUTHORS   Garcia A, Aragones V, Gioiosa S, Herraiz FJ, Ortiz-Garcia P, Prohens
            J, Daros JA, Pasin F
  TITLE     Comparative genomics-driven design of virus-delivered short RNA 
            inserts triggering robust gene silencing.
  JOURNAL   Plant Biotechnol J. 2025 Nov;23(11):4930-4932. doi: 
            10.1111/pbi.70254. Epub 2025 Jul 24.
  PUBMED    41170645
REFERENCE   2  (bases 1 to 5853)
  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"; doi: "10.1111/pbi.70254";
            journalName: "Plant Biotechnol J"; date: "2025-11"; volume: "23"; 
            issue: "11"; pages: "4930-4932"
COMMENT     Sequence Label: pLX-TRV2-vCHLI
FEATURES             Location/Qualifiers
     source          1..5853
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    133..157
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    184..208
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     507..527
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        516..562
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             complement(3391..4206)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     4198..4217
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(4332..4994)
                     /codon_start=1
                     /product="replication protein for the broad-host-range 
                     plasmid pBBR1 from Bordetella bronchiseptica"
                     /label=pBBR1 Rep
                     /translation="MATQSREIGIQAKNKPGHWVQTERKAHEAWAGLIARKPTAAMLLH
                     HLVAQMGHQNAVVVSQKTLSKLIGRSLRTVQYAVKDLVAERWISVVKLNGPGTVSAYVV
                     NDRVAWGQPRDQLRLSVFSAAVVVDHDDQDESLLGHGDLRRIPTLYPGEQQLPTGPGEE
                     PPSQPGIPGMEPDLPALTETEEWERRGQQRLPMPDEPCFLDDGEPLEPPTRVTLPRR"
     rep_origin      4995..5766
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
ORIGIN
        1 ccatcatcag ttcggtggtc ttccgacgaa caataaggcc gcaaatcgcg gcctttttta
       61 ttgataacaa aaccggctca gttctgcgta gaaaccaaca tgcaagctcc accgggtgca
      121 aagcggcagc ggcggcagga tatattcaat tgtaaatggc ttcatgtccg ggaaatctac
      181 tggtggcagg atatattgtg gtgtaaacaa tggagaaaaa gattaattaa gtactagtag
      241 cctggtcgaa accgtctcag attccattgc ccagctatct gtcactttat tgtgaagata
      301 gtggaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa ggccatcgtt
      361 gaagatgcct ctgccgacag tggtcccaaa gatggacccc cacccacgag gagcatcgtg
      421 gaaaaagaag acgttccaac cacgtcttca aagcaagtgg attgatgtga tatctccact
      481 gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc tatataagga
      541 agttcatttc atttggagag gataaaacat tgcacctatg gtgttgccct ggctggggta
      601 tgtcagtgat cgcagtagaa tgtactaatt gacaagttgg agaatacggt agaacgtcct
      661 tatccaacac agcctttatc cctctccctg acgaggtttt tgtcagtgta atatttcttt
      721 ttgaactatc cagcttagta ccgtacggga aagtgactgg tgtgcttatc tttgaaatgt
      781 tactttgggt ttcggttctt taggttagta agaaagcact tgtcttctca tacaaaggaa
      841 aacctgagac gtatcgctta cgaaagtagc aatgaaagaa aggtggtggt tttaatcgct
      901 accgcaaaaa cgatggggtc gttttaatta acttctccta cgcaagcgtc taaacggacg
      961 ttggggtttt gctagtttct ttagagaaaa ctagctaagt ctttaatgtt atcattagag
     1021 atggcataaa tataatactt gtgtctgctg ataagatcat tttaatttgg acgattagac
     1081 ttgttgaact acaggttact gaatcacttg cgctaatcaa catgggagat atgtacgatg
     1141 aatcatttga caagtcgggc ggtcctgctg acttgatgga cgattcttgg gtggaatcag
     1201 tttcgtggaa agatctgttg aagaagttac acagcataaa atttgcacta cagtctggta
     1261 gagatgagat cactgggtta ctagcggcac tgaatagaca gtgtccttat tcaccatatg
     1321 agcagtttcc agataagaag gtgtatttcc ttttagactc acgggctaac agtgctcttg
     1381 gtgtgattca gaacgcttca gcgttcaaga gacgagctga tgagaagaat gcagtggcgg
     1441 gtgttacaaa tattcctgcg aatccaaaca caacggttac gacgaaccaa gggagtacta
     1501 ctactaccaa ggcgaacact ggctcgactt tggaagaaga cttgtacact tattacaaat
     1561 tcgatgatgc ctctacagct ttccacaaat ctctaacttc gttagagaac atggagttga
     1621 agagttatta ccgaaggaac tttgagaaag tattcgggat taagtttggt ggagcagctg
     1681 ctagttcatc tgcaccgcct ccagcgagtg gaggtccgat acgtcctaat ccctagggat
     1741 ttaaggacgt gaactctgtt gagatctctg tgaaattcag agggtgggtg ataccatatt
     1801 cactgatgcc attagcgaca tctaaatagg gctaattgtg actaatttga gggaatttcc
     1861 tttaccattg acgtcagtgt cgttggtagc atttgagttt cgcaatgcac gaattactta
     1921 ggaagtggct tgacgacact aatgtgttat tgttagataa tggtttggtg gtcaaggtac
     1981 gtagtagagt cccacatatt cgcacgtatg aagtaattgg aaagttgtca gtttttgata
     2041 attcactggg agatgatacg ctgtttgagg gaaaagtaga gaacgtattt gtttttatgt
     2101 tcaggcggtt cttgtgtgtc aacaaagatg gacattgtta ctcaaggaag cacgatgagc
     2161 tttattatta cggacgagtg gacttagatt ctgtgagtaa ggttaccgaa ttctgcacac
     2221 aaggttaaaa acgctgtagt aatacatgcg caagaacagg ctgagcatct tgttctgggg
     2281 tttcacacta tctttagaga aagtgttaag ttaattaagt tatcttaatt aagagcataa
     2341 ttatactgat ttgtctctcg ttgatagagt ctatcattct gttactaaaa atttgacaac
     2401 tcggtttgct gacctactgg ttactgtatc acttacccga gttaacgcct gaaacttggg
     2461 catgcattcc aaatcgatca agaagctcta tgtcccgaag acattaaact acggttcttt
     2521 aagtagatcc gtgtctgaag ttttaggttc aatttaaacc tacgagattg acattctcga
     2581 ctgatcttga ttgatcggta agtcttttgt aatttaattt tctttttgat tttattttaa
     2641 attgttatct gtttctgtgt atagactgtt tgagatcggc gtttggccga ctcattgtct
     2701 taccataggg gaacggactt tgtttgtgtt gttattttat ttgtatttta ttaaaattct
     2761 caacgatctg aaaaagcctc gcggctaaga gattgttggg gggtgagtaa gtacttttaa
     2821 agtgatgatg gttacaaagg caaaaggggt aaaacccctc gcctacgtaa gcgttattac
     2881 gcccgggtcg gcatggcatc tccacctcct cgcggtccga cctgggctac ttcggtaggc
     2941 taagggagaa gcgctgaaat caccagtctc tctctacaaa tctatctctc tctattttct
     3001 ccataaataa tgtgtgagta gtttcccgat aagggaaatt agggttctta tagggtttcg
     3061 ctcatgtgtt gagcatataa gaaaccctta gtatgtattt gtatttgtaa aatacttcta
     3121 tcaataaaat ttctaattcc taaaaccaaa attcaggtca ggagacggga caaggatgcg
     3181 agctagcctg caggaattgt tgattttgtg atgactgatg gcaggatata tgcggttgta
     3241 attcattttt attgtctaaa tttctgtatt tgtttgtttg ttcggttgta aatttttttg
     3301 gaagaacaag aaaagaaaaa acacccgtta gggtgttttt agttagtgtg gcgcgccgac
     3361 ttgcgacatg cggtcctttg caatcaacta ttagaaaaat tcatccagca tcagatgaaa
     3421 ttgcagtttg ttcatatccg gattatcaat gccatatttc tgaaacagac gtttttgcag
     3481 gctcgggcta aattcgccca ggcagttcca cagaatggcc agatcctgat aacgatccgc
     3541 aatgcccaca cggcccacat caatgcagcc aatcagtttg ccttcatcga aaatcaggtt
     3601 atccaggcta aaatcgccgt gggtcaccac gctatccggg ctaaacggca gcagtttatg
     3661 catttctttc cacacctgtt ccaccggcca gccgttacgt tcatcatcaa aatcgctcgc
     3721 atccaccagg ccgttgttca tacggctctg cgcctgggcc agacgaaaca cacgatcgct
     3781 gttaaacggg cagttgcaca ccggaatgct atgcagacga cgcagaaaca cggccagcgc
     3841 atccacaatg ttttcgccgc tatccggata ttcttccagc acctgaaacg cggttttgcc
     3901 cggaatcgcg gtggtcagca gccacgcatc atccggggtg cgaataaaat gtttaatggt
     3961 cggcagcggc ataaattcgg tcagccagtt cagacgcacc atttcatcgg tcacatcgtt
     4021 cgccacgctg cctttgccat gtttcagaaa cagttccggc gcatccggtt tgccatacag
     4081 acgataaatg gtcgcgccgc tctgacccac gttatcacgc gcccatttat agccatacag
     4141 atccgcatcc atgttgctgt tcagacgcgg acggctacag ctcgtttcac gctgaatatg
     4201 gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg ttcatgatga
     4261 tatattttta tcttgtgcaa tgtaacatca gagattttga gacacaagat cggattggcg
     4321 gttatgcggt tctaccggcg cggcagcgtt acccgtgtcg gcggctccaa cggctcgcca
     4381 tcgtccagaa aacacggctc atcgggcatc ggcaggcgct gctgcccgcg ccgttcccat
     4441 tcctccgttt cggtcaaggc tggcaggtct ggttccatgc ccggaatgcc gggctggctg
     4501 ggcggctcct cgccggggcc ggtcggtagt tgctgctcgc ccggatacag ggtcgggatg
     4561 cggcgcaggt cgccatgccc caacagcgat tcgtcctggt cgtcgtgatc aaccaccacg
     4621 gcggcactga acaccgacag gcgcaactgg tcgcggggct ggccccacgc cacgcggtca
     4681 ttgaccacgt aggccgacac ggtgccgggg ccgttgagct tcacgacgga gatccagcgc
     4741 tcggccacca agtccttgac tgcgtattgg accgtccgca aagaacgtcc gatgagcttg
     4801 gaaagtgtct tctggctgac caccacggcg ttctggtggc ccatctgcgc cacgaggtga
     4861 tgcagcagca ttgccgccgt gggtttcctc gcaataagcc cggcccacgc ctcatgcgct
     4921 ttgcgttccg tttgcaccca gtgaccgggc ttgttcttgg cttgaatgcc gatttctctg
     4981 gactgcgtgg ccatgcttat ctccatgcgg taggggtgcc gcacggttgc ggcaccatgc
     5041 gcaatcagct gcaacttttc ggcagcgcga caacaattat gcgttgcgta aaagtggcag
     5101 tcaattacag attttcttta acctacgcaa tgagctattg cggggggtgc cgcaatgagc
     5161 tgttgcgtac cccccttttt taagttgttg atttttaagt ctttcgcatt tcgccctata
     5221 tctagttctt tggtgcccaa agaagggcac ccctgcgggg ttcccccacg ccttcggcgc
     5281 ggctccccct ccggcaaaaa gtggcccctc cggggcttgt tgatcgactg cgcggccttc
     5341 ggccttgccc aaggtggcgc tgcccccttg gaacccccgc actcgccgcc gtgaggctcg
     5401 gggggcaggc gggcgggctt cgcccttcga ctgcccccac tcgcataggc ttgggtcgtt
     5461 ccaggcgcgt caaggccaag ccgctgcgcg gtcgctgcgc gagccttgac ccgccttcca
     5521 cttggtgtcc aaccggcaag cgaagcgcgc aggccgcagg ccggaggctt ttccccagag
     5581 aaaattaaaa aaattgatgg ggcaaggccg caggccgcgc agttggagcc ggtgggtatg
     5641 tggtcgaagg ctgggtagcc ggtgggcaat ccctgtggtc aagctcgtgg gcaggcgcag
     5701 cctgtccatc agcttgtcca gcagggttgt ccacgggccg agcgaagcga gccagccggt
     5761 ggccgctcgc ggccatcgtc cacatatcca cgggctggca agggagcgca gcgaccgcgc
     5821 agggcgaagc ccggagagca agcccgtagg ggg
//
