LOCUS       sequence-355071-        6241 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Agrobacterium tumefaciens-based expression of TRV2 including a PEBV 
            heterologous promoter and apolylinker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6241)
  AUTHORS   Aragones V, Aliaga F, Pasin F, Daros JA
  TITLE     Simplifying plant gene silencing and genome editing logistics by a 
            one-Agrobacterium system for simultaneous delivery of multipartite 
            virus vectors.
  JOURNAL   Biotechnol J. 2022 Mar 24:e2100504. doi: 10.1002/biot.202100504.
  PUBMED    35332696
REFERENCE   2  (bases 1 to 6241)
  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: "Biotechnol 
            J. 2022 Mar 24:e2100504. doi: 10.1002/biot.202100504."
COMMENT     Sequence Label: pLX-TRV2
FEATURES             Location/Qualifiers
     source          1..6241
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     248..268
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     promoter        257..303
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     protein_bind    2233..2254
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2269..2274,2275..2292,2293..2299)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2307..2323
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2312..2334
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2331..2347
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2331..2347
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2343..2636
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWVPNSPYSESYYARSLAVVLQRRDWEN
                     PGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAGGSRSMSRRH"
     primer_bind     2366..2386
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2368..2386
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2412..2431)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2413..2431)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2441..2458)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2441..2457)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2450..2472)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             complement(3520..4335)
                     /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     4327..4346
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             complement(4461..5123)
                     /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      5124..5895
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid 
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     misc_feature    6115..6139
                     /label=LB T-DNA repeat
                     /note="left border repeat from octopine Ach5 T-DNA"
     misc_feature    6166..6190
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 gattccattg cccagctatc tgtcacttta ttgtgaagat agtggaaaag gaaggtggct
       61 cctacaaatg ccatcattgc gataaaggaa aggccatcgt tgaagatgcc tctgccgaca
      121 gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa
      181 ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac
      241 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
      301 ggataaaaca ttgcacctat ggtgttgccc tggctggggt atgtcagtga tcgcagtaga
      361 atgtactaat tgacaagttg gagaatacgg tagaacgtcc ttatccaaca cagcctttat
      421 ccctctccct gacgaggttt ttgtcagtgt aatatttctt tttgaactat ccagcttagt
      481 accgtacggg aaagtgactg gtgtgcttat ctttgaaatg ttactttggg tttcggttct
      541 ttaggttagt aagaaagcac ttgtcttctc atacaaagga aaacctgaga cgtatcgctt
      601 acgaaagtag caatgaaaga aaggtggtgg ttttaatcgc taccgcaaaa acgatggggt
      661 cgttttaatt aacttctcct acgcaagcgt ctaaacggac gttggggttt tgctagtttc
      721 tttagagaaa actagctaag tctttaatgt tatcattaga gatggcataa atataatact
      781 tgtgtctgct gataagatca ttttaatttg gacgattaga cttgttgaac tacaggttac
      841 tgaatcactt gcgctaatca acatgggaga tatgtacgat gaatcatttg acaagtcggg
      901 cggtcctgct gacttgatgg acgattcttg ggtggaatca gtttcgtgga aagatctgtt
      961 gaagaagtta cacagcataa aatttgcact acagtctggt agagatgaga tcactgggtt
     1021 actagcggca ctgaatagac agtgtcctta ttcaccatat gagcagtttc cagataagaa
     1081 ggtgtatttc cttttagact cacgggctaa cagtgctctt ggtgtgattc agaacgcttc
     1141 agcgttcaag agacgagctg atgagaagaa tgcagtggcg ggtgttacaa atattcctgc
     1201 gaatccaaac acaacggtta cgacgaacca agggagtact actactacca aggcgaacac
     1261 tggctcgact ttggaagaag acttgtacac ttattacaaa ttcgatgatg cctctacagc
     1321 tttccacaaa tctctaactt cgttagagaa catggagttg aagagttatt accgaaggaa
     1381 ctttgagaaa gtattcggga ttaagtttgg tggagcagct gctagttcat ctgcaccgcc
     1441 tccagcgagt ggaggtccga tacgtcctaa tccctaggga tttaaggacg tgaactctgt
     1501 tgagatctct gtgaaattca gagggtgggt gataccatat tcactgatgc cattagcgac
     1561 atctaaatag ggctaattgt gactaatttg agggaatttc ctttaccatt gacgtcagtg
     1621 tcgttggtag catttgagtt tcgcaatgca cgaattactt aggaagtggc ttgacgacac
     1681 taatgtgtta ttgttagata atggtttggt ggtcaaggta cgtagtagag tcccacatat
     1741 tcgcacgtat gaagtaattg gaaagttgtc agtttttgat aattcactgg gagatgatac
     1801 gctgtttgag ggaaaagtag agaacgtatt tgtttttatg ttcaggcggt tcttgtgtgt
     1861 caacaaagat ggacattgtt actcaaggaa gcacgatgag ctttattatt acggacgagt
     1921 ggacttagat tctgtgagta aggttaccga attctgcaca caaggttaaa aacgctgtag
     1981 taatacatgc gcaagaacag gctgagcatc ttgttctggg gtttcacact atctttagag
     2041 aaagtgttaa gttaattaag ttatcttaat taagagcata attatactga tttgtctctc
     2101 gttgatagag tctatcattc tgttactaaa aatttgacaa ctcggtttgc tgacctactg
     2161 gttactgtat cacttacccg agttaacgcc tgaaggagac cgcgggaaag cgggcagtga
     2221 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
     2281 gctcccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
     2341 ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac aaaagctggg
     2401 tacccaattc gccctatagt gagtcgtatt acgcgcgctc actggccgtc gttttacaac
     2461 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt
     2521 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca
     2581 gcctgaatgg cgaatgggac gcgggcgggt ctcgctctat gtcccgaaga cattaaacta
     2641 cggttcttta agtagatccg tgtctgaagt tttaggttca atttaaacct acgagattga
     2701 cattctcgac tgatcttgat tgatcggtaa gtcttttgta atttaatttt ctttttgatt
     2761 ttattttaaa ttgttatctg tttctgtgta tagactgttt gagatcggcg tttggccgac
     2821 tcattgtctt accatagggg aacggacttt gtttgtgttg ttattttatt tgtattttat
     2881 taaaattctc aacgatctga aaaagcctcg cggctaagag attgttgggg ggtgagtaag
     2941 tacttttaaa gtgatgatgg ttacaaaggc aaaaggggta aaacccctcg cctacgtaag
     3001 cgttattacg cccgggtcgg catggcatct ccacctcctc gcggtccgac ctgggctact
     3061 tcggtaggct aagggagaag cgctgaaatc accagtctct ctctacaaat ctatctctct
     3121 ctattttctc cataaataat gtgtgagtag tttcccgata agggaaatta gggttcttat
     3181 agggtttcgc tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa
     3241 atacttctat caataaaatt tctaattcct aaaaccaaaa ttcaggtcag gagacgggac
     3301 aaggatgcga gctagcctgc aggaattgtt gattttgtga tgactgatgg caggatatat
     3361 gcggttgtaa ttcattttta ttgtctaaat ttctgtattt gtttgtttgt tcggttgtaa
     3421 atttttttgg aagaacaaga aaagaaaaaa cacccgttag ggtgttttta gttagtgtgg
     3481 cgcgccgact tgcgacatgc ggtcctttgc aatcaactat tagaaaaatt catccagcat
     3541 cagatgaaat tgcagtttgt tcatatccgg attatcaatg ccatatttct gaaacagacg
     3601 tttttgcagg ctcgggctaa attcgcccag gcagttccac agaatggcca gatcctgata
     3661 acgatccgca atgcccacac ggcccacatc aatgcagcca atcagtttgc cttcatcgaa
     3721 aatcaggtta tccaggctaa aatcgccgtg ggtcaccacg ctatccgggc taaacggcag
     3781 cagtttatgc atttctttcc acacctgttc caccggccag ccgttacgtt catcatcaaa
     3841 atcgctcgca tccaccaggc cgttgttcat acggctctgc gcctgggcca gacgaaacac
     3901 acgatcgctg ttaaacgggc agttgcacac cggaatgcta tgcagacgac gcagaaacac
     3961 ggccagcgca tccacaatgt tttcgccgct atccggatat tcttccagca cctgaaacgc
     4021 ggttttgccc ggaatcgcgg tggtcagcag ccacgcatca tccggggtgc gaataaaatg
     4081 tttaatggtc ggcagcggca taaattcggt cagccagttc agacgcacca tttcatcggt
     4141 cacatcgttc gccacgctgc ctttgccatg tttcagaaac agttccggcg catccggttt
     4201 gccatacaga cgataaatgg tcgcgccgct ctgacccacg ttatcacgcg cccatttata
     4261 gccatacaga tccgcatcca tgttgctgtt cagacgcgga cggctacagc tcgtttcacg
     4321 ctgaatatgg ctcataacac cccttgtatt actgtttatg taagcagaca gttttattgt
     4381 tcatgatgat atatttttat cttgtgcaat gtaacatcag agattttgag acacaagatc
     4441 ggattggcgg ttatgcggtt ctaccggcgc ggcagcgtta cccgtgtcgg cggctccaac
     4501 ggctcgccat cgtccagaaa acacggctca tcgggcatcg gcaggcgctg ctgcccgcgc
     4561 cgttcccatt cctccgtttc ggtcaaggct ggcaggtctg gttccatgcc cggaatgccg
     4621 ggctggctgg gcggctcctc gccggggccg gtcggtagtt gctgctcgcc cggatacagg
     4681 gtcgggatgc ggcgcaggtc gccatgcccc aacagcgatt cgtcctggtc gtcgtgatca
     4741 accaccacgg cggcactgaa caccgacagg cgcaactggt cgcggggctg gccccacgcc
     4801 acgcggtcat tgaccacgta ggccgacacg gtgccggggc cgttgagctt cacgacggag
     4861 atccagcgct cggccaccaa gtccttgact gcgtattgga ccgtccgcaa agaacgtccg
     4921 atgagcttgg aaagtgtctt ctggctgacc accacggcgt tctggtggcc catctgcgcc
     4981 acgaggtgat gcagcagcat tgccgccgtg ggtttcctcg caataagccc ggcccacgcc
     5041 tcatgcgctt tgcgttccgt ttgcacccag tgaccgggct tgttcttggc ttgaatgccg
     5101 atttctctgg actgcgtggc catgcttatc tccatgcggt aggggtgccg cacggttgcg
     5161 gcaccatgcg caatcagctg caacttttcg gcagcgcgac aacaattatg cgttgcgtaa
     5221 aagtggcagt caattacaga ttttctttaa cctacgcaat gagctattgc ggggggtgcc
     5281 gcaatgagct gttgcgtacc cccctttttt aagttgttga tttttaagtc tttcgcattt
     5341 cgccctatat ctagttcttt ggtgcccaaa gaagggcacc cctgcggggt tcccccacgc
     5401 cttcggcgcg gctccccctc cggcaaaaag tggcccctcc ggggcttgtt gatcgactgc
     5461 gcggccttcg gccttgccca aggtggcgct gcccccttgg aacccccgca ctcgccgccg
     5521 tgaggctcgg ggggcaggcg ggcgggcttc gcccttcgac tgcccccact cgcataggct
     5581 tgggtcgttc caggcgcgtc aaggccaagc cgctgcgcgg tcgctgcgcg agccttgacc
     5641 cgccttccac ttggtgtcca accggcaagc gaagcgcgca ggccgcaggc cggaggcttt
     5701 tccccagaga aaattaaaaa aattgatggg gcaaggccgc aggccgcgca gttggagccg
     5761 gtgggtatgt ggtcgaaggc tgggtagccg gtgggcaatc cctgtggtca agctcgtggg
     5821 caggcgcagc ctgtccatca gcttgtccag cagggttgtc cacgggccga gcgaagcgag
     5881 ccagccggtg gccgctcgcg gccatcgtcc acatatccac gggctggcaa gggagcgcag
     5941 cgaccgcgca gggcgaagcc cggagagcaa gcccgtaggg ggccatcatc agttcggtgg
     6001 tcttccgacg aacaataagg ccgcaaatcg cggccttttt tattgataac aaaaccggct
     6061 cagttctgcg tagaaaccaa catgcaagct ccaccgggtg caaagcggca gcggcggcag
     6121 gatatattca attgtaaatg gcttcatgtc cgggaaatct actggtggca ggatatattg
     6181 tggtgtaaac aatggagaaa aagattaatt aagtactagt agcctggtcg aaaccgtctc
     6241 a
//
