LOCUS       sequence-455257-        7406 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Level 1 - Position 1 with 35S promoter and terminator for plant 
            expression, with mutations in CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7406)
  TITLE     Biosynthesis of anti-inflammatory triterpene fatty acid esters in 
            Calendula officinalis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7406)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pEPHS1CB0030
FEATURES             Location/Qualifiers
     source          1..7406
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(157..261)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      348..979
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    1071..1095
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        1281..1626
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     1569..1589
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    4060..4234
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    4279..4303
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4459..5607)
                     /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(5943..6531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6023..6042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6702..7406,1..87,88..156))
                     /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     7325..7344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
       61 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
      121 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
      181 aagcatttat caaggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
      241 taaacaaata ggggttccgc gcacgaattg gccagcgctg ccatttttgg ggtgaggccg
      301 ttcgcggccg aggggcgcag cccctggggg gatgggaggc ccgcgttagc gggccgggag
      361 ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc
      421 ctggttaaaa acaaggttta taaatattgg tttaaaagca ggttaaaaga caggttagcg
      481 gtggccgaaa aacgggcgga aacccttgca aatgctggat tttctgcctg tggacagccc
      541 ctcaaatgtc aataggtgcg cccctcatct gtcagcactc tgcccctcaa gtgtcaagga
      601 tcgcgcccct catctgtcag tagtcgcgcc cctcaagtgt caataccgca gggcacttat
      661 ccccaggctt gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat
      721 ttgcgaggct ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa
      781 cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag
      841 ggccaagttt tccgcgaggt atccacaacg ccggcggccg cggtgtctcg cacacggctt
      901 cgacggcgtt tctggcgcgt ttgcagggcc atagacggcc gccagcccag cggcgagggc
      961 aaccagcccg gtgagcgtcg caaaggagat cctgatctga ctgatgggct gcctgtatcg
     1021 agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata
     1081 tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta
     1141 atgtactggg gtggatgcag tgggccccac tctgtgaaga caatgccgaa ttcggatccg
     1201 gaggtcaaca tggtggagca cgacactctg gtctactcca aaaatgtcaa agatacagtc
     1261 tcagaagatc aaagggctat tgagactttt caacaaagga taatttcggg aaacctcctc
     1321 ggattccatt gcccagctat ctgtcacttc atcgaaagga cagtagaaaa ggaaggtggc
     1381 tcctacaaat gccatcattg cgataaagga aaggctatca ttcaagatct ctctgccgac
     1441 agtggtccca aagatggacc cccacccacg aggagcatcg tggaaaaaga agaggttcca
     1501 accacgtcta caaagcaagt ggattgatgt gacatctcca ctgacgtaag ggatgacgca
     1561 caatcccact atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag
     1621 aggacacgct cgagtataag agctcatttt tacaacaatt accaacaaca acaaacaaca
     1681 aacaacatta caattacatt tacaattatc gatacaatgt ggaagttgaa gatagcagaa
     1741 gggaatgatc cttacttatt tagtaccaac aatttcgtcg gtaggcaaat ttgggaattt
     1801 gatcccgacg ctggtactcc cgaagaaaaa caagaagttg aaaatgctcg tcaacattat
     1861 agaaacaatc gaaaggaaga tgttcatcca tgcagtgatt tgcttatgcg gatgcagttg
     1921 atcaaagaga atggcataga tctactgagt attccaccag caaggttagg agataatgag
     1981 caagtaaatt atgaagcagt gacgagatca gtgacgaaag cagttcgatt gaaccgtgcg
     2041 attcaagcaa aggatggtca ctggcctgca gaaaacgctg gccctatgtt tttcactcct
     2101 cccctcctta ttgctatgta catcagtgga gccatcgcca cacatttaac caaagaacac
     2161 aagaccgaaa tgatacgtta tatctacaac catcaaaacg atgatggagg gtggggattt
     2221 tatatcgagg gacacagtac catgatcggg tctgctttga gctatgtagc cctaaggtta
     2281 ctaggagagg gacctcatga tggaaatggt gcggttgacc gagccagaaa gtggataatt
     2341 gaccatggtg gtgcaacctc aattccctca tggggcaaaa cttatctatc ggttcttgga
     2401 gtatatgaat gggaaggatg caatccactt ccaccagaat tttggatttt ccctgaaact
     2461 ttaccatatc atccggcgaa aatgtggtgc tattgccgga caacatacat gcctatgtca
     2521 tacttatatg ggacaaaatt tcatggtccg atcactgatc ttgttctaca acttagacaa
     2581 gaaattcatg cgatcccgta caatgagata aattggaata aacaacgaca taactgttgt
     2641 aaggaagatt tgtattaccc tcattcaacg ttgcaagatc tattgtggga tggacttcat
     2701 tacttaagtg aaccgtttct taaatattgg ccttttaaaa agctaagaga aagaggactc
     2761 aaaagagcag ttgaattaat gcgatatggt gctcaacaaa gcagatatat gactattgga
     2821 tgtgttgata agagcttgca aatgatgtgt tggtgggcag agaacccaaa tggggatgaa
     2881 ttcaagcacc atcttgctag agtgcctgat tatttgtggt tagcagaaga tggaatgaag
     2941 atgcaaagtt ttgggagcca attatgggat tgcacacttg caacacaagc aataatagct
     3001 actgatatgg ttgaagaata tggtgattca cttaaaaaag ctcattttta tatcaaagaa
     3061 tctcaagtta aacaaaatcc ttcgggcgat tttagtcaaa tgtgcagaca gtttactata
     3121 gggtcgtgga ctttttctga ccaagatcat ggttgggttg tctcagattg cacagctgaa
     3181 gcattaaagt gtcttttgtt actatcccaa atgccagagg aaattgtggg agaaaaagct
     3241 gataatgagc gattgtatga agctgttaat gttcttcttt acttacagag tcctattact
     3301 ggaggttttg ctatttggga gccacctgtc ccgcaaccat atttacaaat gttgaatcct
     3361 tcagaacttt tcgcagatat tgttgttgag aaagagcatg tcgagtgtac agcatcaatt
     3421 attcaagcac ttttagcctt caaacgactg cacccagggc acagggagaa agaaattgaa
     3481 atttccgtgg caaaagcagt tacatttttg gagggaaaac aacagcatga tggttcatgg
     3541 tatggttatt ggggaatatg ctttctatat ggcacattct ttgctttagg aggcttagtt
     3601 tctgctggaa aaacatataa taacagtgaa gaaattcgta aagcagttaa ttttttcatt
     3661 ttgactcaaa atgaagaagg cggttgggga gaaagtatca aatcttgccc tactgaagta
     3721 tacacaccct tggatggaaa ccgaacaaac ctagttcaaa catcatgggc tatgcttgct
     3781 cttatgttag gtggacaggc tgaaagagat ccaacacctt tgcataaagc agcaaagata
     3841 ttaattaatg cacagatgga taatggagat tttcctcaac aagagattac tggagtttac
     3901 atgaagaatt gcatgctaca ttatccagaa tacagaaaca tcttcccact ttgggcactt
     3961 ggcgaatatc gcaaacgagt ttgggtcaat ggttcgtaag tcgaccggct gctaacaaag
     4021 cccgagcttc tctagctaga gtcgatcgac aagctcgagt ttctccataa taatgtgtga
     4081 gtagttccca gataagggaa ttagggttcc tatagggttt cgctcatgtg ttgagcatat
     4141 aagaaaccct tagtatgtat ttgtatttgt aaaatacttc tatcaataaa atttctaatt
     4201 cctaaaacca aaatccagta ctaaaatcca gatcgctgca attgtcttct gcacgaagtg
     4261 gtttaaacta tcagtgtttg acaggatata ttggcgggta aacctaagag aaaagagcgt
     4321 ttattagaat aatcggatat ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt
     4381 atgtgcatgc caaccacagg gttccccaga tcaggcgctg gctgctgaac ccccagccgg
     4441 aactgacccc acaaggccct agcgtttgca atgcaccagg tcatcattga cccaggcgtg
     4501 ttccaccagg ccgctgcctc gcaactcttc gcaggcttcg ccgacctgct cgcgccactt
     4561 cttcacgcgg gtggaatccg atccgcacat gaggcggaag gtttccagct tgagcgggta
     4621 cggctcccgg tgcgagctga aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg
     4681 gtacttctcc catatgaatt tcgtgtagtg gtcgccagca aacagcacga cgatttcctc
     4741 gtcgatcagg acctggcaac gggacgtttt cttgccacgg tccaggacgc ggaagcggtg
     4801 cagcagcgac accgattcca ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc
     4861 ctgtaggcgc gacaggcatt cctcggcctt cgtgtaatac cggccattga tcgaccagcc
     4921 caggtcctgg caaagctcgt agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt
     4981 cgcgtactcc aacacctgct gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc
     5041 ggtgtaggtg atcttcacgt ccttgttgac gtggaaaatg accttgtttt gcagcgcctc
     5101 gcgcgggatt ttcttgttgc gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat
     5161 cgctcgcatc gtgtccggcc acggcgcaat atcgaacaag gaaagctgca tttccttgat
     5221 ctgctgcttc gtgtgtttca gcaacgcggc ctgcttggcc tcgctgacct gttttgccag
     5281 gtcctcgccg gcggtttttc gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat
     5341 cgacttcgcc aaacctgccg cctcctgttc aagacgacgc gaacgctcca cggcggccga
     5401 tggcgcgggc agggcagggg gagccagttg cacgctgtcg cgctcgatct tggccgtagc
     5461 ttgctggacc atcgagccga cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg
     5521 gcttgcgatg gtttcggcat cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta
     5581 tgccttccgg tcaaacgtcc gattcattca ccctccttgc gggattgccc cgactcacgc
     5641 cggggcaatg tgcccttatt cctgatttga cccgcctggt gccttggtgt ccagataatc
     5701 caccttatcg gcaatgaagt cggtcccgta gaccgtctgg ccgtccttct cgtacttggt
     5761 attccgaatc ttgccctgca cgaataccag cgaccccttg cccaaatact tgccgtgggc
     5821 ctcggcctga gagccaaaac acttgatgcg gaagaagtcg gtgcgctcct gcttgtcgcc
     5881 ggcatcgttg cgccacatct aggatctgcc aggaaccgta aaaaggccgc gttgctggcg
     5941 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     6001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     6061 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     6121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     6181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     6241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     6301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     6361 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     6421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     6481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     6541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     6601 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     6661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     6721 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     6781 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     6841 cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     6901 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     6961 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     7021 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     7081 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     7141 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     7201 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     7261 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     7321 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     7381 tcggggcgaa aactctcaag gatctt
//
