LOCUS       sequence-433790-        8236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Silencing of Nb F3'5'H.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8236)
  AUTHORS   Grutzner R, Konig K, Horn C, Engler C, Laub A, Vogt T, Marillonnet S
  TITLE     A transient expression tool box for anthocyanin biosynthesis in 
            Nicotiana benthamiana.
  JOURNAL   Plant Biotechnol J. 2023 Dec 20. doi: 10.1111/pbi.14261.
  PUBMED    38124296
REFERENCE   2  (bases 1 to 8236)
  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: "Plant 
            Biotechnol J. 2023 Dec 20. doi: 10.1111/pbi.14261."
COMMENT     Sequence Label: pAGM81963
FEATURES             Location/Qualifiers
     source          1..8236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      13..265
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1504..1528
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1684..2832)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     rep_origin      complement(3168..3756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3248..3267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3927..4718,4719..4787))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4550..4569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4788..4892)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4979..5610
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    5702..5726
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        6824..7169
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     7112..7132
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
ORIGIN
        1 gcttgtcaag cagatcgttc aaacatttgg caataaagtt tcttaagatt gaatcctgtt
       61 gccggtcttg cgatgattat catataattt ctgttgaatt acgttaagca tgtaataatt
      121 aacatgtaat gcatgacgtt atttatgaga tgggttttta tgattagagt cccgcaatta
      181 tacatttaat acgcgataga aaacaaaata tagcgcgcaa actaggataa attatcgcgc
      241 gcggtgtcat ctatgttact agatcgacgc tactattgtc ttccactggg tgtcaaagga
      301 gtgtcagtta attaaagagt gtcggggccg acactcttaa tggaagtaaa agtgtgggtt
      361 ttgctgcacc gacactctta atttaagagt gtcggggtcc cgatgaaacc gacgctttta
      421 atttaagagt gtgggttttt ccacaccgac actcttatga atgttaccct aaattcccca
      481 atcctatact gtataagagc tcatttttac aacaattacc aacaacaaca aacaacaaac
      541 aacattacaa ttacatttac aattatcgat acaatgtctc ctgaaagaag gcctgctgat
      601 attagaaggg ctactgaggc tgatatgcct gctgtgtgca ctattgtgaa tcattacatt
      661 gagacttcta ctgtgaattt caggactgag cctcaagaac cacaagagtg gactgatgat
      721 cttgtgaggc ttagagagag gtatccttgg cttgttgctg aagtggatgg tgaagtggct
      781 ggtattgctt atgctggtcc ttggaaggct aggaatgctt acgattggac tgctgagtct
      841 actgtttacg tgtctcctag gcatcaaaga actggtcttg gatctactct ttacactcat
      901 cttttgaagt ctttggaggc tcagggtttc aagtctgtgg ttgctgtgat tggacttcct
      961 aatgatcctt ctgtgaggat gcatgaggct cttggttatg ctcctagggg aatgcttaga
     1021 gctgctggtt tcaaacatgg aaattggcat gatgtgggat tctggcagct tgatttctct
     1081 cttcctgttc ctcctagacc tgttcttcct gtgactgaga tttgagctta ctccacctaa
     1141 atactatgcc ggttcaaata agaaagtaat gattatgtac ccaaataata catttataag
     1201 atgtatgtgt gtgatggaca tgtttctgta tcgtaccaaa actaaatagg tggcttttat
     1261 cccaaaaaag tgctttcgtt tgatgacctt atgatttcta aagagtgttg ttgttgctgc
     1321 ttatgtttaa taaaagtgat gtatattatc agttgatttc cccacgtgga aggggtgatt
     1381 atattgcaca aaattaagtt aaaagtaaca ttgtggtgta ataaaaaccc tagtgaattt
     1441 ataacctata agtaagtggt gcagtagtga atttatcacg aagtggttta aactatcagt
     1501 gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg
     1561 gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc
     1621 acagggttcc ccagatcagg cgctggctgc tgaaccccca gccggaactg accccacaag
     1681 gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct
     1741 gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga
     1801 atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga
     1861 gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat
     1921 gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg
     1981 gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga
     2041 ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag
     2101 gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag
     2161 ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac
     2221 ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt
     2281 cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt
     2341 gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc
     2401 cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg
     2461 tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct cgccggcggt
     2521 ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc
     2581 tgccgcctcc tgttcaagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc
     2641 agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga
     2701 gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc
     2761 ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa
     2821 cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg caatgtgccc
     2881 ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct tatcggcaat
     2941 gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc gaatcttgcc
     3001 ctgcacgaat accagcgacc ccttgcccaa atacttgccg tgggcctcgg cctgagagcc
     3061 aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg tcgccggcat cgttgcgcca
     3121 catctaggat ctgccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     3241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     3301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     3361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     3421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3781 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3841 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3901 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3961 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4021 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca
     4081 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4141 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4201 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     4261 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     4321 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     4381 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     4441 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4501 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4561 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4621 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4681 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4741 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4801 caatattatt gaagcattta tcaaggttat tgtctcatga gcggatacat atttgaatgt
     4861 atttagaaaa ataaacaaat aggggttccg cgcacgaatt ggccagcgct gccatttttg
     4921 gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag
     4981 cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac
     5041 agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag
     5101 acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct
     5161 gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca
     5221 agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc
     5281 agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg
     5341 ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc
     5401 tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat
     5461 ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc gcggtgtctc
     5521 gcacacggct tcgacggcgt ttctggcgcg tttgcagggc catagacggc cgccagccca
     5581 gcggcgaggg caaccagccc ggtgagcgtc gcaaaggaga tcctgatctg actgatgggc
     5641 tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg
     5701 gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc
     5761 ggacgttttt aatgtactgg ggtggatgca gtgggcccca ctctgtgaag acaagcaaga
     5821 attcaagctt ggaggaattc caatcccaca aaaatctgag cttaacagca cagttgctcc
     5881 tctcagagca gaatcgggta ttcaacaccc tcatatcaac tactacgttg tgtataacgg
     5941 tccacatgcc ggtatatacg atgactgggg ttgtacaaag gcggcaacaa acggcgttcc
     6001 cggagttgca cacaagaaat ttgccactat tacagaggca agagcagcag ctgacgcgta
     6061 cacaacaagt cagcaaacag acaggttgaa cttcatcccc aaaggagaag ctcaactcaa
     6121 gcccaagagc tttgctaagg ccctaacaag cccaccaaag caaaaagccc actggctcac
     6181 gctaggaacc aaaaggccca gcagtgatcc agccccaaaa gagatctcct ttgccccgga
     6241 gattacaatg gacgatttcc tctatcttta cgatctagga aggaagttcg aaggtgaagg
     6301 tgacgacact atgttcacca ctgataatga gaaggttagc ctcttcaatt tcagaaagaa
     6361 tgctgaccca cagatggtta gagaggccta cgcagcaagt ctcatcaaga cgatctaccc
     6421 gagtaacaat ctccaggaga tcaaatacct tcccaagaag gttaaagatg cagtcaaaag
     6481 attcaggact aattgcatca agaacacaga gaaagacata tttctcaaga tcagaagtac
     6541 tattccagta tggacgattc aaggcttgct tcataaacca aggcaagtaa tagagattgg
     6601 agtctctaaa aaggtagttc ctactgaatc taaggccatg catggagtct aagattcaaa
     6661 tcgaggatct aacagaactc gccgtcaaga ctggcgaaca gttcatacag agtcttttac
     6721 gactcaatga caagaagaaa atcttcgtca acatggtgga gcacgacact ctggtctact
     6781 ccaaaaatgt caaagataca gtctcagaag atcaaagggc tattgagact tttcaacaaa
     6841 ggataatttc gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa
     6901 ggacagtaga aaaggaaggt ggctcctaca aatgccatca ttgcgataaa ggaaaggcta
     6961 tcattcaaga tctctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca
     7021 tcgtggaaaa agaagaggtt ccaaccacgt ctacaaagca agtggattga tgtgacatct
     7081 ccactgacgt aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat
     7141 aaggaagttc atttcatttg gagaggacac gctactgagt ggcttgtgca cgagataatg
     7201 gtggccggtg atcggcgcac taccatactt aggagccatg ccacatgttt cacttgcaaa
     7261 aatggccaaa aaatatggac ctgttatgta tttaaaagtt ggcacatgtg gtatggctgt
     7321 tgcttctact cctgatgctg ctaaagcatt cttgaaaaca cttgatatca acttctccaa
     7381 tcgtccacct aatgcaggtg ccacgcactt ggcctatgat gcacaagaca tggtttttgc
     7441 acattatgga ccacgttgga agttgctaag gaaattgagc aacttgcata tgctaggggg
     7501 aaaagcctta gaaaattggg caaatgtaag gtaaatcccc agatgaacat ggcatcgtgg
     7561 tgattgatga aactgctgct gtcggcttta acctctcttt aggcattggt ttcgaagcgg
     7621 gcaacaagcc gaaagaactg tacagcgaag aggcagtcaa cggggaaact cagcaagcgc
     7681 acttacaggc gattaaagag ctgatagcgc gtgacaaaaa ccacccaagc gtggtgatgt
     7741 ggagtattgc caacgaaccg gatacccgtc cgcaaggtgc acgggaatat ttcgcgccac
     7801 tggcggaagc aacgcgtaaa ctcgacccga cgcgtccgat cacctgcgtc aatgtaatgt
     7861 tctgcgacgc tcacaccgat accatcagcg atctctttga tgggcaggta catttgccca
     7921 attttctaag gcttttcccc ctagcatatg caagttgctc aatttcctta gcaacttcca
     7981 acgtggtcca taatgtgcaa aaaccatgtc ttgtgcatca taggccaagt gcgtggcacc
     8041 tgcattaggt ggacgattgg agaagttgat atcaagtgtt ttcaagaatg ctttagcagc
     8101 atcaggagta gaagcaacag ccataccaca tgtgccaact tttaaataca taacaggtcc
     8161 atattttttg gccatttttg caagtgaaac atgtggcatg gctcctaagt atggtagtgc
     8221 gccgatcacc ggccac
//
