LOCUS       sequence-455223-        6170 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 6170)
  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 6170)
  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: pEPDG1CB0001
FEATURES             Location/Qualifiers
     source          1..6170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    135..159
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        345..690
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     633..653
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    1888..2062
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2107..2131
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2287..3435)
                     /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(3771..4359)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3851..3870)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4530..5321,5322..5390))
                     /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     5153..5172
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5391..5495)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5582..6170,1..43)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 ggccgccagc ccagcggcga gggcaaccag cccggtgagc gtcgcaaagg agatcctgat
       61 ctgactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc
      121 tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt
      181 aataacacat tgcggacgtt tttaatgtac tggggtggat gcagtgggcc ccactctgtg
      241 aagacaatgc cgaattcgga tccggaggtc aacatggtgg agcacgacac tctggtctac
      301 tccaaaaatg tcaaagatac agtctcagaa gatcaaaggg ctattgagac ttttcaacaa
      361 aggataattt cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcgaa
      421 aggacagtag aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggct
      481 atcattcaag atctctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc
      541 atcgtggaaa aagaagaggt tccaaccacg tctacaaagc aagtggattg atgtgacatc
      601 tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata
      661 taaggaagtt catttcattt ggagaggaca cgctcgagta taagagctca tttttacaac
      721 aattaccaac aacaacaaac aacaaacaac attacaatta catttacaat tatcgataca
      781 atgatgaata acctcatcaa gatatcattt tatgcatata tttcacttgg gtattgcttt
      841 cttgttggca agattgtccc aaaaggtgtc ccaagactcc ttagtatcct ccctgtcgtc
      901 atcatcttcc tctctatccc tctatcttta acatccgttc actgcattgg agccgtttct
      961 ttcttcattt catggcttgc aaacttcaaa ctcatccttt tcgccttcga aaaaggcccc
     1021 ctttcatcac cttctatatc acttatgtgg ttctttgcaa tggcttgctt cccaatccaa
     1081 gtttcacggt ccaataaagg gttttcaacc ccgaaaacac ctagaaaaaa agtgtttagt
     1141 tatggtacaa agggcgtact tcttaccctt ttccttcggg tttattataa ccatggtgcg
     1201 gttatgaacc cgatgctagc atggtgtttc ttagggttct cgctttacct aatgctcgag
     1261 ctcttgctca tactcagctc agggattgta ctattgtttc tcacgattga gctagacccc
     1321 caatttgacg agccttatct ttccacgtcc ctacaagact tttggggacg aaggtggaac
     1381 attatggtca accggatcct tcatccgacc atttacgacc cgattcttaa actatcagat
     1441 cgggtcgtgg gtcggttgtg ggctccggtt ccagccattt tgaccacctt cgcagtttct
     1501 gggctcatgc atgagctcat tttcttttac ttcacccgag cttggcctac ttgggaggtg
     1561 atgcttttct tttgtctaca tggggtttgt ttggtcacgg aggtaatgtt taagaaactt
     1621 cttaacgtta aatggagtct accacgtcaa ttaacagcac cgttagtcgt catattcgtg
     1681 ttggcaacgt cgtgttggtt gtttctcccg gaattactaa ggtgtaaaat gcttgaaagg
     1741 gcgtttcaag agtacgaggc ggttagcaag ttagcaatgg atgttattca aagtccccta
     1801 atatttttga agtataatgg ttcgtaagtc gaccggctgc taacaaagcc cgagcttctc
     1861 tagctagagt cgatcgacaa gctcgagttt ctccataata atgtgtgagt agttcccaga
     1921 taagggaatt agggttccta tagggtttcg ctcatgtgtt gagcatataa gaaaccctta
     1981 gtatgtattt gtatttgtaa aatacttcta tcaataaaat ttctaattcc taaaaccaaa
     2041 atccagtact aaaatccaga tcgctgcaat tgtcttctgc acgaagtggt ttaaactatc
     2101 agtgtttgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt attagaataa
     2161 tcggatattt aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca
     2221 accacagggt tccccagatc aggcgctggc tgctgaaccc ccagccggaa ctgaccccac
     2281 aaggccctag cgtttgcaat gcaccaggtc atcattgacc caggcgtgtt ccaccaggcc
     2341 gctgcctcgc aactcttcgc aggcttcgcc gacctgctcg cgccacttct tcacgcgggt
     2401 ggaatccgat ccgcacatga ggcggaaggt ttccagcttg agcgggtacg gctcccggtg
     2461 cgagctgaaa tagtcgaaca tccgtcgggc cgtcggcgac agcttgcggt acttctccca
     2521 tatgaatttc gtgtagtggt cgccagcaaa cagcacgacg atttcctcgt cgatcaggac
     2581 ctggcaacgg gacgttttct tgccacggtc caggacgcgg aagcggtgca gcagcgacac
     2641 cgattccagg tgcccaacgc ggtcggacgt gaagcccatc gccgtcgcct gtaggcgcga
     2701 caggcattcc tcggccttcg tgtaataccg gccattgatc gaccagccca ggtcctggca
     2761 aagctcgtag aacgtgaagg tgatcggctc gccgataggg gtgcgcttcg cgtactccaa
     2821 cacctgctgc cacaccagtt cgtcatcgtc ggcccgcagc tcgacgccgg tgtaggtgat
     2881 cttcacgtcc ttgttgacgt ggaaaatgac cttgttttgc agcgcctcgc gcgggatttt
     2941 cttgttgcgc gtggtgaaca gggcagagcg ggccgtgtcg tttggcatcg ctcgcatcgt
     3001 gtccggccac ggcgcaatat cgaacaagga aagctgcatt tccttgatct gctgcttcgt
     3061 gtgtttcagc aacgcggcct gcttggcctc gctgacctgt tttgccaggt cctcgccggc
     3121 ggtttttcgc ttcttggtcg tcatagttcc tcgcgtgtcg atggtcatcg acttcgccaa
     3181 acctgccgcc tcctgttcaa gacgacgcga acgctccacg gcggccgatg gcgcgggcag
     3241 ggcaggggga gccagttgca cgctgtcgcg ctcgatcttg gccgtagctt gctggaccat
     3301 cgagccgacg gactggaagg tttcgcgggg cgcacgcatg acggtgcggc ttgcgatggt
     3361 ttcggcatcc tcggcggaaa accccgcgtc gatcagttct tgcctgtatg ccttccggtc
     3421 aaacgtccga ttcattcacc ctccttgcgg gattgccccg actcacgccg gggcaatgtg
     3481 cccttattcc tgatttgacc cgcctggtgc cttggtgtcc agataatcca ccttatcggc
     3541 aatgaagtcg gtcccgtaga ccgtctggcc gtccttctcg tacttggtat tccgaatctt
     3601 gccctgcacg aataccagcg accccttgcc caaatacttg ccgtgggcct cggcctgaga
     3661 gccaaaacac ttgatgcgga agaagtcggt gcgctcctgc ttgtcgccgg catcgttgcg
     3721 ccacatctag gatctgccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3781 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3841 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3901 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3961 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4021 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4081 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4141 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4201 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4261 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4321 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4381 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     4441 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     4501 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     4561 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     4621 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agaaccacgc
     4681 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     4741 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     4801 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4861 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4921 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4981 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5041 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5101 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5161 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5221 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5281 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5341 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     5401 tttcaatatt attgaagcat ttatcaaggt tattgtctca tgagcggata catatttgaa
     5461 tgtatttaga aaaataaaca aataggggtt ccgcgcacga attggccagc gctgccattt
     5521 ttggggtgag gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt
     5581 tagcgggccg ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg
     5641 cacagggcgc agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa
     5701 aagacaggtt agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg
     5761 cctgtggaca gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc
     5821 tcaagtgtca aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac
     5881 cgcagggcac ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag
     5941 gcgttttcgc cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc
     6001 ccctcatctg tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct
     6061 catctgtcag tgagggccaa gttttccgcg aggtatccac aacgccggcg gccgcggtgt
     6121 ctcgcacacg gcttcgacgg cgtttctggc gcgtttgcag ggccatagac
//
