LOCUS       sequence-433793-        7199 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Silencing of Nb RT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7199)
  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 7199)
  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: pAGM81987
FEATURES             Location/Qualifiers
     source          1..7199
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(31..376)
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     complement(68..88)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    1474..1498
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      1590..2221
                     /label=oriV
                     /note="incP origin of replication"
     promoter        2308..2412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2413..2481,2482..3273)
                     /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     complement(2631..2650)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3444..4032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3933..3952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             4368..5516
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    5672..5696
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(6935..7187)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 attatctcgt gcacaagcca ctcagtagcg tgtcctctcc aaatgaaatg aacttcctta
       61 tatagaggaa gggtcttgcg aaggatagtg ggattgtgcg tcatccctta cgtcagtgga
      121 gatgtcacat caatccactt gctttgtaga cgtggttgga acctcttctt tttccacgat
      181 gctcctcgtg ggtgggggtc catctttggg accactgtcg gcagagagat cttgaatgat
      241 agcctttcct ttatcgcaat gatggcattt gtaggagcca ccttcctttt ctactgtcct
      301 ttcgatgaag tgacagatag ctgggcaatg gaatccgagg aggtttcccg aaattatcct
      361 ttgttgaaaa gtctcaatag ccctttgatc ttctgagact gtatctttga catttttgga
      421 gtagaccaga gtgtcgtgct ccaccatgtt gacgaagatt ttcttcttgt cattgagtcg
      481 taaaagactc tgtatgaact gttcgccagt cttgacggcg agttctgtta gatcctcgat
      541 ttgaatctta gactccatgc atggccttag attcagtagg aactaccttt ttagagactc
      601 caatctctat tacttgcctt ggtttatgaa gcaagccttg aatcgtccat actggaatag
      661 tacttctgat cttgagaaat atgtctttct ctgtgttctt gatgcaatta gtcctgaatc
      721 ttttgactgc atctttaacc ttcttgggaa ggtatttgat ctcctggaga ttgttactcg
      781 ggtagatcgt cttgatgaga cttgctgcgt aggcctctct aaccatctgt gggtcagcat
      841 tctttctgaa attgaagagg ctaaccttct cattatcagt ggtgaacata gtgtcgtcac
      901 cttcaccttc gaacttcctt cctagatcgt aaagatagag gaaatcgtcc attgtaatct
      961 ccggggcaaa ggagatctct tttggggctg gatcactgct gggccttttg gttcctagcg
     1021 tgagccagtg ggctttttgc tttggtgggc ttgttagggc cttagcaaag ctcttgggct
     1081 tgagttgagc ttctcctttg gggatgaagt tcaacctgtc tgtttgctga cttgttgtgt
     1141 acgcgtcagc tgctgctctt gcctctgtaa tagtggcaaa tttcttgtgt gcaactccgg
     1201 gaacgccgtt tgttgccgcc tttgtacaac cccagtcatc gtatataccg gcatgtggac
     1261 cgttatacac aacgtagtag ttgatatgag ggtgttgaat acccgattct gctctgagag
     1321 gagcaactgt gctgttaagc tcagattttt gtgggattgg aattcctcca agcttgaatt
     1381 cttgcttgtc ttcacagagt ggggcccact gcatccaccc cagtacatta aaaacgtccg
     1441 caatgtgtta ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc
     1501 agccagccaa cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag
     1561 cccatcagtc agatcaggat ctcctttgcg acgctcaccg ggctggttgc cctcgccgct
     1621 gggctggcgg ccgtctatgg ccctgcaaac gcgccagaaa cgccgtcgaa gccgtgtgcg
     1681 agacaccgcg gccgccggcg ttgtggatac ctcgcggaaa acttggccct cactgacaga
     1741 tgaggggcgg acgttgacac ttgaggggcc gactcacccg gcgcggcgtt gacagatgag
     1801 gggcaggctc gatttcggcc ggcgacgtgg agctggccag cctcgcaaat cggcgaaaac
     1861 gcctgatttt acgcgagttt cccacagatg atgtggacaa gcctggggat aagtgccctg
     1921 cggtattgac acttgagggg cgcgactact gacagatgag gggcgcgatc cttgacactt
     1981 gaggggcaga gtgctgacag atgaggggcg cacctattga catttgaggg gctgtccaca
     2041 ggcagaaaat ccagcatttg caagggtttc cgcccgtttt tcggccaccg ctaacctgtc
     2101 ttttaacctg cttttaaacc aatatttata aaccttgttt ttaaccaggg ctgcgccctg
     2161 tgcgcgtgac cgcgcacgcc gaaggggggt gccccccctt ctcgaaccct cccggcccgc
     2221 taacgcgggc ctcccatccc cccaggggct gcgcccctcg gccgcgaacg gcctcacccc
     2281 aaaaatggca gcgctggcca attcgtgcgc ggaaccccta tttgtttatt tttctaaata
     2341 cattcaaata tgtatccgct catgagacaa taaccttgat aaatgcttca ataatattga
     2401 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     2461 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     2521 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2581 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2641 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2701 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2761 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2821 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2881 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2941 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3001 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3061 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3121 gagcgtggtt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3181 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3241 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     3301 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     3361 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     3421 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     3481 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     3541 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     3601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     3661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     3721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     3781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     3841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     3901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     3961 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     4021 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcaga tcctagatgt
     4081 ggcgcaacga tgccggcgac aagcaggagc gcaccgactt cttccgcatc aagtgttttg
     4141 gctctcaggc cgaggcccac ggcaagtatt tgggcaaggg gtcgctggta ttcgtgcagg
     4201 gcaagattcg gaataccaag tacgagaagg acggccagac ggtctacggg accgacttca
     4261 ttgccgataa ggtggattat ctggacacca aggcaccagg cgggtcaaat caggaataag
     4321 ggcacattgc cccggcgtga gtcggggcaa tcccgcaagg agggtgaatg aatcggacgt
     4381 ttgaccggaa ggcatacagg caagaactga tcgacgcggg gttttccgcc gaggatgccg
     4441 aaaccatcgc aagccgcacc gtcatgcgtg cgccccgcga aaccttccag tccgtcggct
     4501 cgatggtcca gcaagctacg gccaagatcg agcgcgacag cgtgcaactg gctccccctg
     4561 ccctgcccgc gccatcggcc gccgtggagc gttcgcgtcg tcttgaacag gaggcggcag
     4621 gtttggcgaa gtcgatgacc atcgacacgc gaggaactat gacgaccaag aagcgaaaaa
     4681 ccgccggcga ggacctggca aaacaggtca gcgaggccaa gcaggccgcg ttgctgaaac
     4741 acacgaagca gcagatcaag gaaatgcagc tttccttgtt cgatattgcg ccgtggccgg
     4801 acacgatgcg agcgatgcca aacgacacgg cccgctctgc cctgttcacc acgcgcaaca
     4861 agaaaatccc gcgcgaggcg ctgcaaaaca aggtcatttt ccacgtcaac aaggacgtga
     4921 agatcaccta caccggcgtc gagctgcggg ccgacgatga cgaactggtg tggcagcagg
     4981 tgttggagta cgcgaagcgc acccctatcg gcgagccgat caccttcacg ttctacgagc
     5041 tttgccagga cctgggctgg tcgatcaatg gccggtatta cacgaaggcc gaggaatgcc
     5101 tgtcgcgcct acaggcgacg gcgatgggct tcacgtccga ccgcgttggg cacctggaat
     5161 cggtgtcgct gctgcaccgc ttccgcgtcc tggaccgtgg caagaaaacg tcccgttgcc
     5221 aggtcctgat cgacgaggaa atcgtcgtgc tgtttgctgg cgaccactac acgaaattca
     5281 tatgggagaa gtaccgcaag ctgtcgccga cggcccgacg gatgttcgac tatttcagct
     5341 cgcaccggga gccgtacccg ctcaagctgg aaaccttccg cctcatgtgc ggatcggatt
     5401 ccacccgcgt gaagaagtgg cgcgagcagg tcggcgaagc ctgcgaagag ttgcgaggca
     5461 gcggcctggt ggaacacgcc tgggtcaatg atgacctggt gcattgcaaa cgctagggcc
     5521 ttgtggggtc agttccggct gggggttcag cagccagcgc ctgatctggg gaaccctgtg
     5581 gttggcatgc acatacaaat ggacgaacgg ataaaccttt tcacgccctt ttaaatatcc
     5641 gattattcta ataaacgctc ttttctctta ggtttacccg ccaatatatc ctgtcaaaca
     5701 ctgatagttt aaaccacttc gtgataaatt cactactgca ccacttactt ataggttata
     5761 aattcactag ggtttttatt acaccacaat gttactttta acttaatttt gtgcaatata
     5821 atcacccctt ccacgtgggg aaatcaactg ataatataca tcacttttat taaacataag
     5881 cagcaacaac aacactcttt agaaatcata aggtcatcaa acgaaagcac ttttttggga
     5941 taaaagccac ctatttagtt ttggtacgat acagaaacat gtccatcaca cacatacatc
     6001 ttataaatgt attatttggg tacataatca ttactttctt atttgaaccg gcatagtatt
     6061 taggtggagt aagctcaaat ctcagtcaca ggaagaacag gtctaggagg aacaggaaga
     6121 gagaaatcaa gctgccagaa tcccacatca tgccaatttc catgtttgaa accagcagct
     6181 ctaagcattc ccctaggagc ataaccaaga gcctcatgca tcctcacaga aggatcatta
     6241 ggaagtccaa tcacagcaac cacagacttg aaaccctgag cctccaaaga cttcaaaaga
     6301 tgagtgtaaa gagtagatcc aagaccagtt ctttgatgcc taggagacac gtaaacagta
     6361 gactcagcag tccaatcgta agcattccta gccttccaag gaccagcata agcaatacca
     6421 gccacttcac catccacttc agcaacaagc caaggatacc tctctctaag cctcacaaga
     6481 tcatcagtcc actcttgtgg ttcttgaggc tcagtcctga aattcacagt agaagtctca
     6541 atgtaatgat tcacaatagt gcacacagca ggcatatcag cctcagtagc ccttctaata
     6601 tcagcaggcc ttctttcagg agacattgta tcgataattg taaatgtaat tgtaatgttg
     6661 tttgttgttt gttgttgttg gtaattgttg taaaaatgag ctcttataca gtataggatt
     6721 ggggaattta gggtaacatt cataagagtg tcggtgtgga aaaacccaca ctcttaaatt
     6781 aaaagcgtcg gtttcatcgg gaccccgaca ctcttaaatt aagagtgtcg gtgcagcaaa
     6841 acccacactt ttacttccat taagagtgtc ggccccgaca ctctttaatt aactgacact
     6901 cctttgacac ccagtggaag acaatagtag cgtcgatcta gtaacataga tgacaccgcg
     6961 cgcgataatt tatcctagtt tgcgcgctat attttgtttt ctatcgcgta ttaaatgtat
     7021 aattgcggga ctctaatcat aaaaacccat ctcataaata acgtcatgca ttacatgtta
     7081 attattacat gcttaacgta attcaacaga aattatatga taatcatcgc aagaccggca
     7141 acaggattca atcttaagaa actttattgc caaatgtttg aacgatctgc ttgacaagc
//
