LOCUS       sequence-455459-        6569 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 6569)
  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 6569)
  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: pEPDG1CB0005
FEATURES             Location/Qualifiers
     source          1..6569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    40..64
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(220..1368)
                     /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(1704..2292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1784..1803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2463..3254,3255..3323))
                     /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     3086..3105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3324..3428)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3515..4146
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    4238..4262
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        4448..4793
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     4736..4756
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    6390..6564
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
ORIGIN
        1 aattgtcttc tgcacgaagt ggtttaaact atcagtgttt gacaggatat attggcgggt
       61 aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg
      121 tttatccgtt cgtccatttg tatgtgcatg ccaaccacag ggttccccag atcaggcgct
      181 ggctgctgaa cccccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag
      241 gtcatcattg acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc
      301 gccgacctgc tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa
      361 ggtttccagc ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg
      421 ggccgtcggc gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc
      481 aaacagcacg acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg
      541 gtccaggacg cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga
      601 cgtgaagccc atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata
      661 ccggccattg atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg
      721 ctcgccgata ggggtgcgct tcgcgtactc caacacctgc tgccacacca gttcgtcatc
      781 gtcggcccgc agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat
      841 gaccttgttt tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga
      901 gcgggccgtg tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa
      961 ggaaagctgc atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc
     1021 ctcgctgacc tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt
     1081 tcctcgcgtg tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt caagacgacg
     1141 cgaacgctcc acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc
     1201 gcgctcgatc ttggccgtag cttgctggac catcgagccg acggactgga aggtttcgcg
     1261 gggcgcacgc atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg aaaaccccgc
     1321 gtcgatcagt tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg
     1381 cgggattgcc ccgactcacg ccggggcaat gtgcccttat tcctgatttg acccgcctgg
     1441 tgccttggtg tccagataat ccaccttatc ggcaatgaag tcggtcccgt agaccgtctg
     1501 gccgtccttc tcgtacttgg tattccgaat cttgccctgc acgaatacca gcgacccctt
     1561 gcccaaatac ttgccgtggg cctcggcctg agagccaaaa cacttgatgc ggaagaagtc
     1621 ggtgcgctcc tgcttgtcgc cggcatcgtt gcgccacatc taggatctgc caggaaccgt
     1681 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     1741 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     1801 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     1861 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     1921 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     1981 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2041 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2101 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2161 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2221 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2281 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     2341 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     2401 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     2461 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     2521 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     2581 cccagtgctg caatgatacc gcgagaacca cgctcaccgg ctccagattt atcagcaata
     2641 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     2701 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     2761 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     2821 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     2881 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     2941 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     3001 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     3061 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     3121 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     3181 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     3241 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     3301 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcaa
     3361 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3421 gttccgcgca cgaattggcc agcgctgcca tttttggggt gaggccgttc gcggccgagg
     3481 ggcgcagccc ctggggggat gggaggcccg cgttagcggg ccgggagggt tcgagaaggg
     3541 ggggcacccc ccttcggcgt gcgcggtcac gcgcacaggg cgcagccctg gttaaaaaca
     3601 aggtttataa atattggttt aaaagcaggt taaaagacag gttagcggtg gccgaaaaac
     3661 gggcggaaac ccttgcaaat gctggatttt ctgcctgtgg acagcccctc aaatgtcaat
     3721 aggtgcgccc ctcatctgtc agcactctgc ccctcaagtg tcaaggatcg cgcccctcat
     3781 ctgtcagtag tcgcgcccct caagtgtcaa taccgcaggg cacttatccc caggcttgtc
     3841 cacatcatct gtgggaaact cgcgtaaaat caggcgtttt cgccgatttg cgaggctggc
     3901 cagctccacg tcgccggccg aaatcgagcc tgcccctcat ctgtcaacgc cgcgccgggt
     3961 gagtcggccc ctcaagtgtc aacgtccgcc cctcatctgt cagtgagggc caagttttcc
     4021 gcgaggtatc cacaacgccg gcggccgcgg tgtctcgcac acggcttcga cggcgtttct
     4081 ggcgcgtttg cagggccata gacggccgcc agcccagcgg cgagggcaac cagcccggtg
     4141 agcgtcgcaa aggagatcct gatctgactg atgggctgcc tgtatcgagt ggtgattttg
     4201 tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa
     4261 acaaattgac gcttagacaa cttaataaca cattgcggac gtttttaatg tactggggtg
     4321 gatgcagtgg gccccactct gtgaagacaa tgccgaattc ggatccggag gtcaacatgg
     4381 tggagcacga cactctggtc tactccaaaa atgtcaaaga tacagtctca gaagatcaaa
     4441 gggctattga gacttttcaa caaaggataa tttcgggaaa cctcctcgga ttccattgcc
     4501 cagctatctg tcacttcatc gaaaggacag tagaaaagga aggtggctcc tacaaatgcc
     4561 atcattgcga taaaggaaag gctatcattc aagatctctc tgccgacagt ggtcccaaag
     4621 atggaccccc acccacgagg agcatcgtgg aaaaagaaga ggttccaacc acgtctacaa
     4681 agcaagtgga ttgatgtgac atctccactg acgtaaggga tgacgcacaa tcccactatc
     4741 cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg acacgctcga
     4801 gtataagagc tcatttttac aacaattacc aacaacaaca aacaacaaac aacattacaa
     4861 ttacatttac aattatcgat acaatggaga tattctatgc ttctcttatc tctctttttg
     4921 ttcttgttgt ttctttttct cttcattttt tcttctacaa ttcaaaacaa cccgaagatc
     4981 agaaactact tccaccgggt agaacgggct ggcccataat cggagaaagt ctcgagtttc
     5041 tcggaagcgg ttggtacggg caccccgaaa agttcatact tgaccgaatg tcaaagtttt
     5101 cgcctatagt atttaggtca tctctcttgc tagaggatgc tgcagtgatg tgtggtgctc
     5161 aagggaacaa attcttgttc tcaaatgaga acaaactagt ccaagtatgg tggcccgctt
     5221 cggttgacaa aatctttccg tcggctaaca aaacctacaa aaccgaaacc attaaaatga
     5281 gaaaaatgct cccgaatttc ctcaaacccg aagccttgca acgatatatt ccaatcatgg
     5341 atattaccac acaaaaacac tttgccgcgg gatgggaagg gaaaaaagaa gttgtgactt
     5401 atgaactcac aagacacttt gctttttggc ttgcgtgcaa gatctttcta gatatcgatg
     5461 atcctgaacg ggtcagatat ttatctggcc cgtttgagag catcgttacg gggatcctgt
     5521 cgatccccat tgatcttccg gggacaccgt ttagaagagg gatcaatggg tccaacttca
     5581 taataaagga acttgtggat attgtgaagc aaaggaaaat tgatttggct caagggaaag
     5641 caacaccaac ccaagatata ttgtcacata tgattctatt tagcgacgaa gatggcgagt
     5701 tcatgaccga acatgacatt tcgggcaaga ttcttgcgtt gcttattggt gggcatgaca
     5761 ctgcgagctc tgctacggct tttattgtta agtatcttgc cgaattgcca gaaatctatg
     5821 atggagtcta caacgaacaa atggaaattg caaaatataa agcagctggg gaattattga
     5881 attgggagga tctatcaaag atgaaatact catggaaagt tgcatgtgaa gttcttagat
     5941 tagtcccacc ggcccaaggt gcttttagag aagcccttac agatttcatg tacaatggtt
     6001 actctatccc caagggctgg aagttatatt ggagtgcaag ctcaacccat aaaaaccccg
     6061 aggttttccc cgaacccgaa aaatttgacc cgtcaagatt tgatggcagc gggccagcac
     6121 cttacacgtt cgtgccattt ggaggaggcc cgcacatgtg tcccggaaaa gagtatgccc
     6181 gattagaaat actcgtgttt atgcatcatc ttgtaacaaa gttcaaatgg gagaaaatta
     6241 ttcccgatga gcccgttgtt ttcaacccga tgcctagccc tgccaaggga cttcctattc
     6301 gtatttaccc tcataaagct ggttcgtaag tcgaccggct gctaacaaag cccgagcttc
     6361 tctagctaga gtcgatcgac aagctcgagt ttctccataa taatgtgtga gtagttccca
     6421 gataagggaa ttagggttcc tatagggttt cgctcatgtg ttgagcatat aagaaaccct
     6481 tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca
     6541 aaatccagta ctaaaatcca gatcgctgc
//
