LOCUS       sequence-455228-        6185 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 - Position 1 with 35S promoter and terminator for plant 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6185)
  AUTHORS   Golubova D, Salmon M, Su H, Tansley C, Kaithakottil GG, Linsmith G, 
            Schudoma C, Swarbreck D, O'Connell MA, Patron NJ
  TITLE     Biosynthesis and bioactivity of anti-inflammatory triterpenoids in 
            Calendula officinalis.
  JOURNAL   Nat Commun. 2025 Jul 28;16(1):6941. doi: 10.1038/s41467-025-62269-w.
  PUBMED    40721615
REFERENCE   2  (bases 1 to 6185)
  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: "Nat 
            Commun."; date: "2025-07-28"; volume: "16(1):6941. doi"; pages: " 
            10.1038/s41467-025-62269-w"
COMMENT     Sequence Label: pEPDG1CB0003
FEATURES             Location/Qualifiers
     source          1..6185
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        109..454
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     397..417
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    1667..1841
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    1886..1910
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2066..3214)
                     /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(3550..4138)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3630..3649)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4309..5100,5101..5169))
                     /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     4932..4951
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5170..5274)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5361..5992
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6084..6108
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 tgtgaagaca atgccgaatt cggatccgga ggtcaacatg gtggagcacg acactctggt
       61 ctactccaaa aatgtcaaag atacagtctc agaagatcaa agggctattg agacttttca
      121 acaaaggata atttcgggaa acctcctcgg attccattgc ccagctatct gtcacttcat
      181 cgaaaggaca gtagaaaagg aaggtggctc ctacaaatgc catcattgcg ataaaggaaa
      241 ggctatcatt caagatctct ctgccgacag tggtcccaaa gatggacccc cacccacgag
      301 gagcatcgtg gaaaaagaag aggttccaac cacgtctaca aagcaagtgg attgatgtga
      361 catctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag acccttcctc
      421 tatataagga agttcatttc atttggagag gacacgctcg agtataagag ctcattttta
      481 caacaattac caacaacaac aaacaacaaa caacattaca attacattta caattatcga
      541 tacaatggag ggagagatga aaaacatgat atttgtatgg gcaacagcgt ttggatctct
      601 cttcatctgc cacaaaatca ccaaagttac tgcaccaggt acaagtcgct tatgcgcatt
      661 catccctgtg gtttgcatct tcttatacct ccccctccgg cttacaactg ttcatttcgc
      721 cggcacaacc tctttcttca tctcttggct tgcaaacttc aaattgattt tgcttgcttt
      781 caataaaggc cctctgtttt caaatccacc ccttcctttc tccaaattca tcctcaccgc
      841 ttgtgttccc attaaaattt catctcatag atctgattct gaacccaaaa aatcaccgac
      901 ggcgatcaaa tactccctca agtttatggt gtttcttctg ttgcttaaag tgtatgaata
      961 cgacgatcaa tttcatcctc tcatcagaat gactctgttt tgtgttcatt tatatgtggc
     1021 tctagaccta ggtttagcca tggttgcata cctggctcga tccatcactg gattcgaact
     1081 cgaaccacag ttcgatgaac cttacctggc tacatctcta caagactttt ggggaaagag
     1141 gtggaatcta atggtcacgg gtatactaca ccctgccgtg tacctacccg tccgagcaat
     1201 ttccggtagg tttctgacca aaaagcgggc cgcacttcct gctcttatgt tttcgtttat
     1261 cgtatccggg ctgatgcacg agttgatttt ttattacatt gggcgattga agcccacatg
     1321 ggaagtcacg gggttcttcg tcattcatgg cgtgctggtg agtatggagg tggtggtaaa
     1381 aagggcggta gatggaagat tccatctgcc agctgcggtt tccaggccgc tggcgttggg
     1441 tttggtgatc gccaccagct tctggttatt ctttccgccg ttcctgcggt ttgattcgga
     1501 agtccgatta tgtgaagaat ttctcgcctt catggaactt gtccgtcgtg gacggtttgt
     1561 tggcccaaag gacgtctttt gcccttactt tagacgcggt tcgtaagtcg accggctgct
     1621 aacaaagccc gagcttctct agctagagtc gatcgacaag ctcgagtttc tccataataa
     1681 tgtgtgagta gttcccagat aagggaatta gggttcctat agggtttcgc tcatgtgttg
     1741 agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat caataaaatt
     1801 tctaattcct aaaaccaaaa tccagtacta aaatccagat cgctgcaatt gtcttctgca
     1861 cgaagtggtt taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa
     1921 agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc
     1981 catttgtatg tgcatgccaa ccacagggtt ccccagatca ggcgctggct gctgaacccc
     2041 cagccggaac tgaccccaca aggccctagc gtttgcaatg caccaggtca tcattgaccc
     2101 aggcgtgttc caccaggccg ctgcctcgca actcttcgca ggcttcgccg acctgctcgc
     2161 gccacttctt cacgcgggtg gaatccgatc cgcacatgag gcggaaggtt tccagcttga
     2221 gcgggtacgg ctcccggtgc gagctgaaat agtcgaacat ccgtcgggcc gtcggcgaca
     2281 gcttgcggta cttctcccat atgaatttcg tgtagtggtc gccagcaaac agcacgacga
     2341 tttcctcgtc gatcaggacc tggcaacggg acgttttctt gccacggtcc aggacgcgga
     2401 agcggtgcag cagcgacacc gattccaggt gcccaacgcg gtcggacgtg aagcccatcg
     2461 ccgtcgcctg taggcgcgac aggcattcct cggccttcgt gtaataccgg ccattgatcg
     2521 accagcccag gtcctggcaa agctcgtaga acgtgaaggt gatcggctcg ccgatagggg
     2581 tgcgcttcgc gtactccaac acctgctgcc acaccagttc gtcatcgtcg gcccgcagct
     2641 cgacgccggt gtaggtgatc ttcacgtcct tgttgacgtg gaaaatgacc ttgttttgca
     2701 gcgcctcgcg cgggattttc ttgttgcgcg tggtgaacag ggcagagcgg gccgtgtcgt
     2761 ttggcatcgc tcgcatcgtg tccggccacg gcgcaatatc gaacaaggaa agctgcattt
     2821 ccttgatctg ctgcttcgtg tgtttcagca acgcggcctg cttggcctcg ctgacctgtt
     2881 ttgccaggtc ctcgccggcg gtttttcgct tcttggtcgt catagttcct cgcgtgtcga
     2941 tggtcatcga cttcgccaaa cctgccgcct cctgttcaag acgacgcgaa cgctccacgg
     3001 cggccgatgg cgcgggcagg gcagggggag ccagttgcac gctgtcgcgc tcgatcttgg
     3061 ccgtagcttg ctggaccatc gagccgacgg actggaaggt ttcgcggggc gcacgcatga
     3121 cggtgcggct tgcgatggtt tcggcatcct cggcggaaaa ccccgcgtcg atcagttctt
     3181 gcctgtatgc cttccggtca aacgtccgat tcattcaccc tccttgcggg attgccccga
     3241 ctcacgccgg ggcaatgtgc ccttattcct gatttgaccc gcctggtgcc ttggtgtcca
     3301 gataatccac cttatcggca atgaagtcgg tcccgtagac cgtctggccg tccttctcgt
     3361 acttggtatt ccgaatcttg ccctgcacga ataccagcga ccccttgccc aaatacttgc
     3421 cgtgggcctc ggcctgagag ccaaaacact tgatgcggaa gaagtcggtg cgctcctgct
     3481 tgtcgccggc atcgttgcgc cacatctagg atctgccagg aaccgtaaaa aggccgcgtt
     3541 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3601 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3661 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3721 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3781 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3841 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3901 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     3961 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4021 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4081 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4141 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4201 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     4261 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4321 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4381 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4441 gataccgcga gaaccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4501 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4561 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4621 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4681 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4741 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4801 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4861 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4921 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4981 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5041 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     5101 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     5161 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcaaggtt attgtctcat
     5221 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacgaa
     5281 ttggccagcg ctgccatttt tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg
     5341 ggggatggga ggcccgcgtt agcgggccgg gagggttcga gaaggggggg cacccccctt
     5401 cggcgtgcgc ggtcacgcgc acagggcgca gccctggtta aaaacaaggt ttataaatat
     5461 tggtttaaaa gcaggttaaa agacaggtta gcggtggccg aaaaacgggc ggaaaccctt
     5521 gcaaatgctg gattttctgc ctgtggacag cccctcaaat gtcaataggt gcgcccctca
     5581 tctgtcagca ctctgcccct caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc
     5641 gcccctcaag tgtcaatacc gcagggcact tatccccagg cttgtccaca tcatctgtgg
     5701 gaaactcgcg taaaatcagg cgttttcgcc gatttgcgag gctggccagc tccacgtcgc
     5761 cggccgaaat cgagcctgcc cctcatctgt caacgccgcg ccgggtgagt cggcccctca
     5821 agtgtcaacg tccgcccctc atctgtcagt gagggccaag ttttccgcga ggtatccaca
     5881 acgccggcgg ccgcggtgtc tcgcacacgg cttcgacggc gtttctggcg cgtttgcagg
     5941 gccatagacg gccgccagcc cagcggcgag ggcaaccagc ccggtgagcg tcgcaaagga
     6001 gatcctgatc tgactgatgg gctgcctgta tcgagtggtg attttgtgcc gagctgccgg
     6061 tcggggagct gttggctggc tggtggcagg atatattgtg gtgtaaacaa attgacgctt
     6121 agacaactta ataacacatt gcggacgttt ttaatgtact ggggtggatg cagtgggccc
     6181 cactc
//
