LOCUS       sequence-455226-        6167 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 6167)
  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 6167)
  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: pEPDG1CB0002
FEATURES             Location/Qualifiers
     source          1..6167
                     /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    1649..1823
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    1868..1892
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2048..3196)
                     /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(3532..4120)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3612..3631)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4291..5082,5083..5151))
                     /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     4914..4933
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5152..5256)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5343..5974
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6066..6090
                     /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 tacaatgaat aacctcataa agatatcatt ttatgcaaat atttcacttg gatattgctt
      601 tcttgtaggc aagattgtcc caaaaggtgt cccaagactc cttagcatcc tccctgttgt
      661 catcatcttc ctctctatcc ctctctcatt aacctctgtt cactgcattg gaggagtttc
      721 tttcttcatt tcatggcttg caaacttcaa actcatcctt tttgccttcg aaaaaggccc
      781 gctttcatcg ccttctatct cacttatgtg gttctttgca atggcttgtt tcccaatcga
      841 tgttacacga tccaataaag ggttttcaaa cccgaaaaca cctagcaaaa aggtgtttag
      901 ttatggtaca aagggcgtac ttcttaccct tttccttcgg gtttattata atcatggtga
      961 ggttatgaac ccaatgttag catggtgctt tttagggttc tcggtttacc taatgctcga
     1021 gctcttggtc acactcagct cagggattgt actattgttt ctcacggttg agctagaccc
     1081 tcagtttgat gagccttatc tttccacgtc cctgcaagac ttttggggac gaaggtggaa
     1141 cattatggtc aaccggatcc ttcatccgac catttacgac ccgatgctga aactatcaga
     1201 tcgggtcatt ggtcggttgt gggctccggt tccagccatt ttgaccacct tcgcagtttc
     1261 tgggctcatg catgagctaa ttttctttta cttcacccga gcttggccta cttgggaggt
     1321 gatgcttttc ttttgtctac atggggtttg tttggttatg gaggtaatgt ttaagaaact
     1381 tattaacgtt aaatggagtc taccacgtca attaacagca ccgttagtcg tcatattcgt
     1441 gttggcaacg tcctattggc tgtttctccc cgaattactg aggtgtaaaa tgcttgaaag
     1501 agcgtttcaa gagtacgagg cggttagcaa gttagcaatg tatgttattc gaagccccct
     1561 aataatttcg aagtataacg gttcgtaagt cgaccggctg ctaacaaagc ccgagcttct
     1621 ctagctagag tcgatcgaca agctcgagtt tctccataat aatgtgtgag tagttcccag
     1681 ataagggaat tagggttcct atagggtttc gctcatgtgt tgagcatata agaaaccctt
     1741 agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc ctaaaaccaa
     1801 aatccagtac taaaatccag atcgctgcaa ttgtcttctg cacgaagtgg tttaaactat
     1861 cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata
     1921 atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc
     1981 aaccacaggg ttccccagat caggcgctgg ctgctgaacc cccagccgga actgacccca
     2041 caaggcccta gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc
     2101 cgctgcctcg caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg
     2161 tggaatccga tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt
     2221 gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc
     2281 atatgaattt cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga
     2341 cctggcaacg ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca
     2401 ccgattccag gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg
     2461 acaggcattc ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc
     2521 aaagctcgta gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca
     2581 acacctgctg ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga
     2641 tcttcacgtc cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt
     2701 tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg
     2761 tgtccggcca cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg
     2821 tgtgtttcag caacgcggcc tgcttggcct cgctgacctg ttttgccagg tcctcgccgg
     2881 cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca
     2941 aacctgccgc ctcctgttca agacgacgcg aacgctccac ggcggccgat ggcgcgggca
     3001 gggcaggggg agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacca
     3061 tcgagccgac ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg
     3121 tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt
     3181 caaacgtccg attcattcac cctccttgcg ggattgcccc gactcacgcc ggggcaatgt
     3241 gcccttattc ctgatttgac ccgcctggtg ccttggtgtc cagataatcc accttatcgg
     3301 caatgaagtc ggtcccgtag accgtctggc cgtccttctc gtacttggta ttccgaatct
     3361 tgccctgcac gaataccagc gaccccttgc ccaaatactt gccgtgggcc tcggcctgag
     3421 agccaaaaca cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc
     3481 gccacatcta ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     3541 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3601 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3661 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3721 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3781 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3841 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3901 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3961 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4021 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4081 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4141 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4201 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4261 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4321 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4381 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg
     4441 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4501 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     4561 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4621 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4681 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4741 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4801 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4861 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4921 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4981 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5041 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5101 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5161 ttttcaatat tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga
     5221 atgtatttag aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgccatt
     5281 tttggggtga ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg
     5341 ttagcgggcc gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc
     5401 gcacagggcg cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta
     5461 aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct
     5521 gcctgtggac agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc
     5581 ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata
     5641 ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca
     5701 ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg
     5761 cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc
     5821 tcatctgtca gtgagggcca agttttccgc gaggtatcca caacgccggc ggccgcggtg
     5881 tctcgcacac ggcttcgacg gcgtttctgg cgcgtttgca gggccataga cggccgccag
     5941 cccagcggcg agggcaacca gcccggtgag cgtcgcaaag gagatcctga tctgactgat
     6001 gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg
     6061 gctggtggca ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca
     6121 ttgcggacgt ttttaatgta ctggggtgga tgcagtgggc cccactc
//
