LOCUS       sequence-455273-        7460 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 7460)
  TITLE     Biosynthesis of anti-inflammatory triterpene fatty acid esters in 
            Calendula officinalis
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7460)
  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            "Unpublished"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: pEPHS1CB0004
FEATURES             Location/Qualifiers
     source          1..7460
                     /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    4114..4288
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    4333..4357
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4513..5661)
                     /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(5997..6585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6077..6096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6756..7460,1..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     7379..7398
                     /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 ggaaattaaa aattggtgaa
     1741 aaaaatggaa agctggaaat tggtgacgga aatggcgatg aatatttgta tagtaccaac
     1801 aatttcgtag ggagacaaac atgggagttt gatccaagtg caggcacaca ggaagaacgt
     1861 gaccaaatcg aaaggattcg agagcaattc ttgaacaata agaagaaact cgatatccat
     1921 tgttgtggtg acttgctcat gcgaacccag cttatcaagg aaagccagat tgatcttact
     1981 agcgaacttc tagtgagact caaagatgaa gaagatgtta actatgaagc tgtgacaatg
     2041 gcggtaaaaa aagcggtcct cttgaaccgt gcaattcaag cttgggacgg tcattggcca
     2101 gctgaaaatg ccggtcccct tttttttacc cctcctctga taattgcctt gtacattagt
     2161 ggtacagtgg atacaatttt aacaaaagaa caccaaaagg agatgattcg ctacatgtac
     2221 atccatcaga atgaagatgg aggatgggga ttctatatat cggggaaaag cacgatgatt
     2281 ggaagtgcat tgaactatgt ggctctaaga cttcttggag aagcttcacc aactgatgat
     2341 gatgacattg gtgctcttgc tagaggccat aaatggatat ttgatcacgg cggtgctaca
     2401 tccatccctt catggggcaa ggtttatctc gcggtgcttg gggtatatga atgggaaggc
     2461 tgcaatcctc tgcctcctga attttggcta tttccttcct ttttgcctta tcatccagca
     2521 aaaatgtggt gttactgtag aacgacatac atgccgatgt cacacttgta tggtaaaagc
     2581 taccatgggc ctattactga tcttgttttg tctttaagaa aagagattca cacccttcct
     2641 taccaccaga ttaactggaa caaacaacga cataattgtt gtaagttgga tctatactac
     2701 cctcattctt ccatacaaga tatgttatgg gatagtctgc actatttttg tgaaccactt
     2761 gtcaaaaaat ggcccttgaa taagctaaga gagaagggac tcaaaagagt ggttgaccta
     2821 atgcgttatg gatccgaaga aggacgttac ataggcatgg gttgtgttga caaggcttta
     2881 caaatgatgt gtttttatgc cgaggatcca aatgggattg acttcaaaca ccacctggct
     2941 agagtccctg attacttgtg gctggcagaa gatggtatga agatgcaaag tttcggtagt
     3001 cagttatggg attgcactct tgtaactcaa gcaattatgg ccagtaatat ggttgatgag
     3061 tatggagatt cactcaaaaa agcccacttt tacttaaaac agtcacagat taaagagaac
     3121 cctaaaggcg atttcacaaa aatgtgtcgt ttattcacga aaggggcatg gactttctct
     3181 gatcaagacc atggatgggt tgtatcagat tgcactgcag aagctttgat ttgtttattg
     3241 gcattgtcac aaatgccaca agatattgcg ggcgaaaagg ctgaagttga tagattatat
     3301 gatgctgtga atgtcctcct ttacctacaa agtcctgaaa gtggtggttt cgctatttgg
     3361 gagccaccag ttccaaaacc atatttacag atgttgaacc cttcagaact ttttgcagac
     3421 atagtggttg agaaagagca tgttgaatgt actggatcca taatccaagc attgaattca
     3481 ttcaaaaatc tgcacccacg acaccgtgag aaagaaatag aagctgctat cgagaaaggc
     3541 atacgttttt tggaaaacaa acaacaaaat gatggttcat ggtacggtta ttggggtatt
     3601 tgttttctct atggcacatt ttttgtgctg caaggtttag tatcatgcgg gaaaacatat
     3661 gaaaatagtg aaaccgttcg aaaagctgtc aattttttgc tctcaacaca gaattcagaa
     3721 ggtggttggg gagagagttt tgagtcatgc cctcaagaga aattcatacc tttggaagga
     3781 aatagaacaa atttggtgca aacttcatgg gccatgcttg gacttctcta tggtggacag
     3841 gttgaaaggg atgtaacacc gttacacaag gcagcaaaat tgctcattaa tggacaattg
     3901 gataatggag attttcctca acaggaaata acgggagtgt acatgaaaaa ctgcatgtta
     3961 cattatgcag aatataggaa cacttttccg ttatgggcgt taggagaata ccgcaaccgt
     4021 gtttggttgc caaagcaaca agtcggttcg taagtcgacc ggctgctaac aaagcccgag
     4081 cttctctagc tagagtcgat cgacaagctc gagtttctcc ataataatgt gtgagtagtt
     4141 cccagataag ggaattaggg ttcctatagg gtttcgctca tgtgttgagc atataagaaa
     4201 cccttagtat gtatttgtat ttgtaaaata cttctatcaa taaaatttct aattcctaaa
     4261 accaaaatcc agtactaaaa tccagatcgc tgcaattgtc ttctgcacga agtggtttaa
     4321 actatcagtg tttgacagga tatattggcg ggtaaaccta agagaaaaga gcgtttatta
     4381 gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc
     4441 atgccaacca cagggttccc cagatcaggc gctggctgct gaacccccag ccggaactga
     4501 ccccacaagg ccctagcgtt tgcaatgcac caggtcatca ttgacccagg cgtgttccac
     4561 caggccgctg cctcgcaact cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac
     4621 gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc agcttgagcg ggtacggctc
     4681 ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc ggcgacagct tgcggtactt
     4741 ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc acgacgattt cctcgtcgat
     4801 caggacctgg caacgggacg ttttcttgcc acggtccagg acgcggaagc ggtgcagcag
     4861 cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag
     4921 gcgcgacagg cattcctcgg ccttcgtgta ataccggcca ttgatcgacc agcccaggtc
     4981 ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta
     5041 ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta
     5101 ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg
     5161 gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg
     5221 catcgtgtcc ggccacggcg caatatcgaa caaggaaagc tgcatttcct tgatctgctg
     5281 cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg acctgttttg ccaggtcctc
     5341 gccggcggtt tttcgcttct tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt
     5401 cgccaaacct gccgcctcct gttcaagacg acgcgaacgc tccacggcgg ccgatggcgc
     5461 gggcagggca gggggagcca gttgcacgct gtcgcgctcg atcttggccg tagcttgctg
     5521 gaccatcgag ccgacggact ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc
     5581 gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt
     5641 ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt gccccgactc acgccggggc
     5701 aatgtgccct tattcctgat ttgacccgcc tggtgccttg gtgtccagat aatccacctt
     5761 atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc ttctcgtact tggtattccg
     5821 aatcttgccc tgcacgaata ccagcgaccc cttgcccaaa tacttgccgt gggcctcggc
     5881 ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc tcctgcttgt cgccggcatc
     5941 gttgcgccac atctaggatc tgccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     6001 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6061 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6121 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6181 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6241 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6301 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6361 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6421 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6481 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6541 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6601 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6661 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6721 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6781 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6841 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagaa
     6901 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6961 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     7021 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7081 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7141 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7201 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7261 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7321 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7381 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7441 cgaaaactct caaggatctt
//
