LOCUS       sequence-349511-        6663 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 4, transcriptional unit for transient 
            expression of 7-deoxyloganic acid hydroxylase (Cr7-DLH) from 
            Catharanthus roseus driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6663)
  AUTHORS   Dudley QM, Jo S, Guerrero DAS, Chhetry M, Smedley MA, Harwood WA, 
            Sherden NH, O'Connor SE, Caputi L, Patron NJ
  TITLE     Reconstitution of monoterpene indole alkaloid biosynthesis in genome
            engineered Nicotiana benthamiana.
  JOURNAL   Commun Biol. 2022 Sep 10;5(1):949. doi: 10.1038/s42003-022-03904-w.
  PUBMED    36088516
REFERENCE   2  (bases 1 to 6663)
  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: "Commun 
            Biol."; date: "2022-09-10"; volume: "5(1):949. doi"; pages: " 
            10.1038/s42003-022-03904-w"
COMMENT     Sequence Label: pEPQD1CB0767
FEATURES             Location/Qualifiers
     source          1..6663
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    125..149
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        335..680
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     623..643
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2371..2545
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2590..2614
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2770..3918)
                     /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(4254..4842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4334..4353)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5013..5804,5805..5873))
                     /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     5636..5655
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5874..5978)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6065..6663,1..33)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 ccagcggcga gggcaaccag cccggtgagc gtcgcaaagg agatcctgat ctgactgatg
       61 ggctgcctgt atcgagtggt gattttgtgc cgagctgccg gtcggggagc tgttggctgg
      121 ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt aataacacat
      181 tgcggacgtt tttaatgtac tggggtggat gcagtgggcc ccactctgtg aagacaatta
      241 cgaattccca tggggaggtc aacatggtgg agcacgacac tctggtctac tccaaaaatg
      301 tcaaagatac agtctcagaa gatcaaaggg ctattgagac ttttcaacaa aggataattt
      361 cgggaaacct cctcggattc cattgcccag ctatctgtca cttcatcgaa aggacagtag
      421 aaaaggaagg tggctcctac aaatgccatc attgcgataa aggaaaggct atcattcaag
      481 atctctctgc cgacagtggt cccaaagatg gacccccacc cacgaggagc atcgtggaaa
      541 aagaagaggt tccaaccacg tctacaaagc aagtggattg atgtgacatc tccactgacg
      601 taagggatga cgcacaatcc cactatcctt cgcaagaccc ttcctctata taaggaagtt
      661 catttcattt ggagaggaca cgctcgagta taagagctca tttttacaac aattaccaac
      721 aacaacaaac aacaaacaac attacaatta catttacaat tatcgataca atggaattga
      781 acttcaaatc aattattttc ttggtttttg ttagtctaac cctttactgg gtctatagaa
      841 tcttggattg ggtatggttt aagcccaaga agctggaaaa atgcttaaga gaacaagggt
      901 ttaagggaaa tccttacaga ttgttcttgg gagatcagta tgatagtgga aaattgatca
      961 gacaagcctt gactaaacct attggtgttg aagaagatgt taagaaacga attgtacctc
     1021 atatactcaa aactgttggg actcatggca agaagtcttt tatgtgggtt ggaagaatac
     1081 caagggtgaa tatcacagat ccagagctga ttaaagaggt cttaacaaaa tactataagt
     1141 tccagaagaa tcatcatgat cttgacccaa ttaccaaatt gcttttgact ggaattggaa
     1201 gcttagaagg tgacccttgg gctaaacgcc gaaaaatcat caatgctgct tttcactttg
     1261 aaaagttaaa gcttatgctt cctgcattct atttgagttg ccgtgatatg gtgacaaaat
     1321 gggataataa ggtccctgaa ggaggatcag cagaggtgga tgtgtggcat gatattgaaa
     1381 ccttaactgg agatgtaatt tcaagaacat tatttggaag taacttcgag gaagggagaa
     1441 gaatatttga actcatgaaa gaacttactg ctcttactat tgatgtcatt cgctcggttt
     1501 atatccctgg acagaggttt cttccaacta agaggaacaa taggatgaga gcaattgaca
     1561 aagaagttag agtaagaata acagaaatta tcaataagaa aatgaaggta atgaaatcag
     1621 gagaagcagc aagtgctgct gatgattttt tgggaatatt gttggaatgc aatctgaatg
     1681 aaataaagga gcaaggaaat aacaaaagtg ctggaatgac tattggagag attattggtg
     1741 agtgcaaact cttctatttt gctggccagg acactacctc gactttactc gtgtggacaa
     1801 tggtcctctt gtctagattc cccgaatggc aaactcgtgc cagagaagaa gtttttcaag
     1861 tctttggtaa taaaaccccg gactatgatg gtatcagcca tcttaaagtt ataacgatga
     1921 tcttgtacga ggttttgagg ttatatactc cagtggctga actgacaaaa gtggctcatg
     1981 aagctacaca actgggaaaa tacttcattc cagcgggcgt gcagctaatg atgccacaaa
     2041 tcctacttca tcatgaccct gaaatatggg gtgaagatgt aatggagttc aaaccagaaa
     2101 gatttgcaga gggagttctc aaggcaacta agagccaagg ctcttttttt ccatttagct
     2161 tgggaccaag aatgtgcatt ggccaaaact ttgctctatt ggaagccaaa atggcaatgt
     2221 ctctaatctt gcgtagattt tctttcgagc tttctccctc ctatgttcat gctcctttca
     2281 ctctcatcac aatgcaaccc caatatggtg ctcatttgat tctgcacaaa ctctaagctt
     2341 ctctagctag agtcgatcga caagctcgag tttctccata ataatgtgtg agtagttccc
     2401 agataaggga attagggttc ctatagggtt tcgctcatgt gttgagcata taagaaaccc
     2461 ttagtatgta tttgtatttg taaaatactt ctatcaataa aatttctaat tcctaaaacc
     2521 aaaatccagt actaaaatcc agatcgctca gattgtcttc tgcacgaagt ggtttaaact
     2581 atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa
     2641 taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg
     2701 ccaaccacag ggttccccag atcaggcgct ggctgctgaa cccccagccg gaactgaccc
     2761 cacaaggccc tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag
     2821 gccgctgcct cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg
     2881 ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg
     2941 gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc
     3001 ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag
     3061 gacctggcaa cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga
     3121 caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg
     3181 cgacaggcat tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg
     3241 gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc
     3301 caacacctgc tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt
     3361 gatcttcacg tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat
     3421 tttcttgttg cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat
     3481 cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt
     3541 cgtgtgtttc agcaacgcgg cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc
     3601 ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc
     3661 caaacctgcc gcctcctgtt caagacgacg cgaacgctcc acggcggccg atggcgcggg
     3721 cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac
     3781 catcgagccg acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat
     3841 ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg
     3901 gtcaaacgtc cgattcattc accctccttg cgggattgcc ccgactcacg ccggggcaat
     3961 gtgcccttat tcctgatttg acccgcctgg tgccttggtg tccagataat ccaccttatc
     4021 ggcaatgaag tcggtcccgt agaccgtctg gccgtccttc tcgtacttgg tattccgaat
     4081 cttgccctgc acgaatacca gcgacccctt gcccaaatac ttgccgtggg cctcggcctg
     4141 agagccaaaa cacttgatgc ggaagaagtc ggtgcgctcc tgcttgtcgc cggcatcgtt
     4201 gcgccacatc taggatctgc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4261 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4321 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4381 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4441 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4501 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4561 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4621 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4681 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4741 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4801 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4861 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4921 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4981 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     5041 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5101 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca
     5161 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5221 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5281 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5341 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5401 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5461 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5521 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5581 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5641 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5701 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5761 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5821 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5881 ctttttcaat attattgaag catttatcaa ggttattgtc tcatgagcgg atacatattt
     5941 gaatgtattt agaaaaataa acaaataggg gttccgcgca cgaattggcc agcgctgcca
     6001 tttttggggt gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg
     6061 cgttagcggg ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac
     6121 gcgcacaggg cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt
     6181 taaaagacag gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt
     6241 ctgcctgtgg acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc
     6301 ccctcaagtg tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa
     6361 taccgcaggg cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat
     6421 caggcgtttt cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc
     6481 tgcccctcat ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc
     6541 cctcatctgt cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccgcgg
     6601 tgtctcgcac acggcttcga cggcgtttct ggcgcgtttg cagggccata gacggccgcc
     6661 agc
//
