LOCUS       sequence-433789-        8859 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transient expression of At DFR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8859)
  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 8859)
  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: pAGM10440
FEATURES             Location/Qualifiers
     source          1..8859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        994..1339
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1282..1302
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     terminator      2586..3293
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    4533..4557
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4713..5861)
                     /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(6197..6785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6277..6296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6956..7747,7748..7816))
                     /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     7579..7598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7817..7921)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      8008..8639
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    8731..8755
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 ggaggaattc caatcccaca aaaatctgag cttaacagca cagttgctcc tctcagagca
       61 gaatcgggta ttcaacaccc tcatatcaac tactacgttg tgtataacgg tccacatgcc
      121 ggtatatacg atgactgggg ttgtacaaag gcggcaacaa acggcgttcc cggagttgca
      181 cacaagaaat ttgccactat tacagaggca agagcagcag ctgacgcgta cacaacaagt
      241 cagcaaacag acaggttgaa cttcatcccc aaaggagaag ctcaactcaa gcccaagagc
      301 tttgctaagg ccctaacaag cccaccaaag caaaaagccc actggctcac gctaggaacc
      361 aaaaggccca gcagtgatcc agccccaaaa gagatctcct ttgccccgga gattacaatg
      421 gacgatttcc tctatcttta cgatctagga aggaagttcg aaggtgaagg tgacgacact
      481 atgttcacca ctgataatga gaaggttagc ctcttcaatt tcagaaagaa tgctgaccca
      541 cagatggtta gagaggccta cgcagcaagt ctcatcaaga cgatctaccc gagtaacaat
      601 ctccaggaga tcaaatacct tcccaagaag gttaaagatg cagtcaaaag attcaggact
      661 aattgcatca agaacacaga gaaagacata tttctcaaga tcagaagtac tattccagta
      721 tggacgattc aaggcttgct tcataaacca aggcaagtaa tagagattgg agtctctaaa
      781 aaggtagttc ctactgaatc taaggccatg catggagtct aagattcaaa tcgaggatct
      841 aacagaactc gccgtcaaga ctggcgaaca gttcatacag agtcttttac gactcaatga
      901 caagaagaaa atcttcgtca acatggtgga gcacgacact ctggtctact ccaaaaatgt
      961 caaagataca gtctcagaag atcaaagggc tattgagact tttcaacaaa ggataatttc
     1021 gggaaacctc ctcggattcc attgcccagc tatctgtcac ttcatcgaaa ggacagtaga
     1081 aaaggaaggt ggctcctaca aatgccatca ttgcgataaa ggaaaggcta tcattcaaga
     1141 tctctctgcc gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa
     1201 agaagaggtt ccaaccacgt ctacaaagca agtggattga tgtgacatct ccactgacgt
     1261 aagggatgac gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc
     1321 atttcatttg gagaggacac gctcgagtat aagagctcat ttttacaaca attaccaaca
     1381 acaacaaaca acaaacaaca ttacaattac atttacaatt atcgatacaa tggttagtca
     1441 gaaagagaca gtgtgtgtaa ccggcgcttc gggtttcatc ggttcatggc tagtgatgcg
     1501 attactagaa cgtggttact ttgttcgtgc caccgttcga gatcccggta atttgaagaa
     1561 agtacaacat cttcttgatt tgccaaacgc caagacgcta ctcactttat ggaaggctga
     1621 tttatctgag gaaggaagct acgatgatgc cataaacgga tgtgacggtg ttttccacgt
     1681 ggcaacaccc atggattttg aatcaaaaga tcctgagaac gaagtgataa agccgacagt
     1741 gaatggaatg ttggggataa tgaaagcatg tgttaaggca aagaccgtac gaagattcgt
     1801 atttacttca tctgccggaa ccgttaatgt agaagaacat cagaagaatg tctatgatga
     1861 aaatgattgg agtgatcttg agtttatcat gtccaaaaag atgacaggat ggatgtattt
     1921 cgtgtcaaaa acgttagcgg agaaagcagc gtgggatttc gccgaagaga aaggattaga
     1981 tttcattagt attattccaa cattggtggt cggtccattc atcacaacgt ctatgccgcc
     2041 tagccttatc accgcgctct ctcctatcac tcggaacgag gcgcattact cgatcataag
     2101 acaaggacag tatgtgcatt tggacgactt atgcaacgct catatcttct tatacgaaca
     2161 agcagccgcc aagggacgtt atatttgttc ctctcatgat gcaaccattc ttactatctc
     2221 caaatttctc aggccaaaat accccgaata taacgtacct tcaacgtttg aaggtgttga
     2281 tgagaatcta aagagcattg aattcagttc caagaagctg acggacatgg ggtttaactt
     2341 caagtatagt ctcgaggaaa tgtttattga atctattgag acatgtcgtc aaaagggttt
     2401 tctcccggtt tcattatcgt accaatccat atcggagatc aagactaaga atgaaaacat
     2461 tgacgtcaaa accggagatg gtttaaccga tggtatgaag ccatgtaaca agacagaaac
     2521 ggggataacc ggcgagagaa ccgatgctcc catgctagca caacagatgt gtgcctaggc
     2581 ttgtcctgct ttaatgagat atgcgagaag cctatgatcg catgatattt gctttcaatt
     2641 ctgttgtgca cgttgtaaaa aacctgagca tgtgtagctc agatccttac cgccggtttc
     2701 ggttcattct aatgaatata tcacccgtta ctatcgtatt tttatgaata atattctccg
     2761 ttcaatttac tgattgtacc ctactactta tatgtacaat attaaaatga aaacaatata
     2821 ttgtgctgaa taggtttata gcgacatcta tgatagagcg ccacaataac aaacaattgc
     2881 gttttattat tacaaatcca attttaaaaa aagcggcaga accggtcaaa cctaaaagac
     2941 tgattacata aatcttattc aaatttcaaa agtgccccag gggctagtat ctacgacaca
     3001 ccgagcggcg aactaataac gctcactgaa gggaactccg gttccccgcc ggcgcgcatg
     3061 ggtgagattc cttgaagttg agtattggcc gtccgctcta ccgaaagtta cgggcaccat
     3121 tcaacccggt ccagcacggc ggccgggtaa ccgacttgct gccccgagaa ttatgcagca
     3181 tttttttggt gtatgtgggc cccaaatgaa gtgcaggtca aaccttgaca gtgacgacaa
     3241 atcgttgggc gggtccaggg cgaattttgc gacaacatgt cgaggctcag caggaccgct
     3301 gcaattgtct tccactgggt gtcaaaggag tgtcagttaa ttaaagagtg tcggggccga
     3361 cactcttaat ggaagtaaaa gtgtgggttt tgctgcaccg acactcttaa tttaagagtg
     3421 tcggggtccc gatgaaaccg acgcttttaa tttaagagtg tgggtttttc cacaccgaca
     3481 ctcttatgaa tgttacccta aattccccaa tcctatactg tataagagct catttttaca
     3541 acaattacca acaacaacaa acaacaaaca acattacaat tacatttaca attatcgata
     3601 caatgtctcc tgaaagaagg cctgctgata ttagaagggc tactgaggct gatatgcctg
     3661 ctgtgtgcac tattgtgaat cattacattg agacttctac tgtgaatttc aggactgagc
     3721 ctcaagaacc acaagagtgg actgatgatc ttgtgaggct tagagagagg tatccttggc
     3781 ttgttgctga agtggatggt gaagtggctg gtattgctta tgctggtcct tggaaggcta
     3841 ggaatgctta cgattggact gctgagtcta ctgtttacgt gtctcctagg catcaaagaa
     3901 ctggtcttgg atctactctt tacactcatc ttttgaagtc tttggaggct cagggtttca
     3961 agtctgtggt tgctgtgatt ggacttccta atgatccttc tgtgaggatg catgaggctc
     4021 ttggttatgc tcctagggga atgcttagag ctgctggttt caaacatgga aattggcatg
     4081 atgtgggatt ctggcagctt gatttctctc ttcctgttcc tcctagacct gttcttcctg
     4141 tgactgagat ttgagcttac tccacctaaa tactatgccg gttcaaataa gaaagtaatg
     4201 attatgtacc caaataatac atttataaga tgtatgtgtg tgatggacat gtttctgtat
     4261 cgtaccaaaa ctaaataggt ggcttttatc ccaaaaaagt gctttcgttt gatgacctta
     4321 tgatttctaa agagtgttgt tgttgctgct tatgtttaat aaaagtgatg tatattatca
     4381 gttgatttcc ccacgtggaa ggggtgatta tattgcacaa aattaagtta aaagtaacat
     4441 tgtggtgtaa taaaaaccct agtgaattta taacctataa gtaagtggtg cagtagtgaa
     4501 tttatcacga agtggtttaa actatcagtg tttgacagga tatattggcg ggtaaaccta
     4561 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc
     4621 gttcgtccat ttgtatgtgc atgccaacca cagggttccc cagatcaggc gctggctgct
     4681 gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac caggtcatca
     4741 ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc ttcgccgacc
     4801 tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc
     4861 agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc
     4921 ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc
     4981 acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc acggtccagg
     5041 acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag
     5101 cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta ataccggcca
     5161 ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg
     5221 ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc
     5281 cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg
     5341 ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc
     5401 gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa caaggaaagc
     5461 tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg
     5521 acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat agttcctcgc
     5581 gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcaagacg acgcgaacgc
     5641 tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct gtcgcgctcg
     5701 atcttggccg tagcttgctg gaccatcgag ccgacggact ggaaggtttc gcggggcgca
     5761 cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc
     5821 agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt
     5881 gccccgactc acgccggggc aatgtgccct tattcctgat ttgacccgcc tggtgccttg
     5941 gtgtccagat aatccacctt atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc
     6001 ttctcgtact tggtattccg aatcttgccc tgcacgaata ccagcgaccc cttgcccaaa
     6061 tacttgccgt gggcctcggc ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc
     6121 tcctgcttgt cgccggcatc gttgcgccac atctaggatc tgccaggaac cgtaaaaagg
     6181 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6241 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6301 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6361 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6421 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6481 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6541 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6601 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6661 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6721 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6781 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6841 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6901 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6961 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     7021 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     7081 ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc
     7141 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7201 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7261 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7321 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7381 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7441 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7501 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7561 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7621 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7681 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7741 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7801 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat caaggttatt
     7861 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7921 gcacgaattg gccagcgctg ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag
     7981 cccctggggg gatgggaggc ccgcgttagc gggccgggag ggttcgagaa gggggggcac
     8041 cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta
     8101 taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga
     8161 aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg
     8221 cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag
     8281 tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca
     8341 tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc
     8401 acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg
     8461 cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt
     8521 atccacaacg ccggcggccg cggtgtctcg cacacggctt cgacggcgtt tctggcgcgt
     8581 ttgcagggcc atagacggcc gccagcccag cggcgagggc aaccagcccg gtgagcgtcg
     8641 caaaggagat cctgatctga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
     8701 ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaatt
     8761 gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg gtggatgcag
     8821 tgggccccac tctgtgaaga caatgccgaa ttcggatcc
//
