LOCUS       sequence-349513-        6840 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; contains a chloroplast 
            transit peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6840)
  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 6840)
  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: pEPQD1CB0768
FEATURES             Location/Qualifiers
     source          1..6840
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    122..146
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        332..677
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     620..640
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2545..2719
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2764..2788
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2944..4092)
                     /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(4428..5016)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4508..4527)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5187..5978,5979..6047))
                     /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     5810..5829
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6048..6152)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6239..6840,1..30)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gcggcgaggg caaccagccc ggtgagcgtc gcaaaggaga tcctgatctg actgatgggc
       61 tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg
      121 gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc
      181 ggacgttttt aatgtactgg ggtggatgca gtgggcccca ctctgtgaag acaattacga
      241 attcccatgg ggaggtcaac atggtggagc acgacactct ggtctactcc aaaaatgtca
      301 aagatacagt ctcagaagat caaagggcta ttgagacttt tcaacaaagg ataatttcgg
      361 gaaacctcct cggattccat tgcccagcta tctgtcactt catcgaaagg acagtagaaa
      421 aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg aaaggctatc attcaagatc
      481 tctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc gtggaaaaag
      541 aagaggttcc aaccacgtct acaaagcaag tggattgatg tgacatctcc actgacgtaa
      601 gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa ggaagttcat
      661 ttcatttgga gaggacacgc tactgtataa gagctcattt ttacaacaat taccaacaac
      721 aacaaacaac aaacaacatt acaattacat ttacaattat cgataacatg gcttcttcta
      781 tgctttcttc tgctgctgtt gttgctactc gtgctagtgc tgctcaagct agtatggttg
      841 ctccttttac tggacttaag tctgctgctt cttttcctgt tactagaaag caaaacaacc
      901 ttgatattac ttctattgct agtaacggag gaagagttcg agcaatggaa ttgaacttca
      961 aatcaattat tttcttggtt tttgttagtc taacccttta ctgggtctat agaatcttgg
     1021 attgggtatg gtttaagccc aagaagctgg aaaaatgctt aagagaacaa gggtttaagg
     1081 gaaatcctta cagattgttc ttgggagatc agtatgatag tggaaaattg atcagacaag
     1141 ccttgactaa acctattggt gttgaagaag atgttaagaa acgaattgta cctcatatac
     1201 tcaaaactgt tgggactcat ggcaagaagt cttttatgtg ggttggaaga ataccaaggg
     1261 tgaatatcac agatccagag ctgattaaag aggtcttaac aaaatactat aagttccaga
     1321 agaatcatca tgatcttgac ccaattacca aattgctttt gactggaatt ggaagcttag
     1381 aaggtgaccc ttgggctaaa cgccgaaaaa tcatcaatgc tgcttttcac tttgaaaagt
     1441 taaagcttat gcttcctgca ttctatttga gttgccgtga tatggtgaca aaatgggata
     1501 ataaggtccc tgaaggagga tcagcagagg tggatgtgtg gcatgatatt gaaaccttaa
     1561 ctggagatgt aatttcaaga acattatttg gaagtaactt cgaggaaggg agaagaatat
     1621 ttgaactcat gaaagaactt actgctctta ctattgatgt cattcgctcg gtttatatcc
     1681 ctggacagag gtttcttcca actaagagga acaataggat gagagcaatt gacaaagaag
     1741 ttagagtaag aataacagaa attatcaata agaaaatgaa ggtaatgaaa tcaggagaag
     1801 cagcaagtgc tgctgatgat tttttgggaa tattgttgga atgcaatctg aatgaaataa
     1861 aggagcaagg aaataacaaa agtgctggaa tgactattgg agagattatt ggtgagtgca
     1921 aactcttcta ttttgctggc caggacacta cctcgacttt actcgtgtgg acaatggtcc
     1981 tcttgtctag attccccgaa tggcaaactc gtgccagaga agaagttttt caagtctttg
     2041 gtaataaaac cccggactat gatggtatca gccatcttaa agttataacg atgatcttgt
     2101 acgaggtttt gaggttatat actccagtgg ctgaactgac aaaagtggct catgaagcta
     2161 cacaactggg aaaatacttc attccagcgg gcgtgcagct aatgatgcca caaatcctac
     2221 ttcatcatga ccctgaaata tggggtgaag atgtaatgga gttcaaacca gaaagatttg
     2281 cagagggagt tctcaaggca actaagagcc aaggctcttt ttttccattt agcttgggac
     2341 caagaatgtg cattggccaa aactttgctc tattggaagc caaaatggca atgtctctaa
     2401 tcttgcgtag attttctttc gagctttctc cctcctatgt tcatgctcct ttcactctca
     2461 tcacaatgca accccaatat ggtgctcatt tgattctgca caaactctaa gcttctctag
     2521 ctagagtcga tcgacaagct cgagtttctc cataataatg tgtgagtagt tcccagataa
     2581 gggaattagg gttcctatag ggtttcgctc atgtgttgag catataagaa acccttagta
     2641 tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa aaccaaaatc
     2701 cagtactaaa atccagatcg ctcagattgt cttctgcacg aagtggttta aactatcagt
     2761 gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg
     2821 gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc
     2881 acagggttcc ccagatcagg cgctggctgc tgaaccccca gccggaactg accccacaag
     2941 gccctagcgt ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct
     3001 gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga
     3061 atccgatccg cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga
     3121 gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat
     3181 gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg
     3241 gcaacgggac gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga
     3301 ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag
     3361 gcattcctcg gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag
     3421 ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac
     3481 ctgctgccac accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt
     3541 cacgtccttg ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt
     3601 gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc
     3661 cggccacggc gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg
     3721 tttcagcaac gcggcctgct tggcctcgct gacctgtttt gccaggtcct cgccggcggt
     3781 ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc
     3841 tgccgcctcc tgttcaagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc
     3901 agggggagcc agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga
     3961 gccgacggac tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc
     4021 ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa
     4081 cgtccgattc attcaccctc cttgcgggat tgccccgact cacgccgggg caatgtgccc
     4141 ttattcctga tttgacccgc ctggtgcctt ggtgtccaga taatccacct tatcggcaat
     4201 gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac ttggtattcc gaatcttgcc
     4261 ctgcacgaat accagcgacc ccttgcccaa atacttgccg tgggcctcgg cctgagagcc
     4321 aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg tcgccggcat cgttgcgcca
     4381 catctaggat ctgccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     4441 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4501 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4561 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4621 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4681 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4741 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4801 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4861 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4921 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4981 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5041 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5101 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5161 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     5221 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     5281 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca
     5341 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5401 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5461 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5521 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5581 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5641 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5701 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5761 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5821 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5881 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5941 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6001 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6061 caatattatt gaagcattta tcaaggttat tgtctcatga gcggatacat atttgaatgt
     6121 atttagaaaa ataaacaaat aggggttccg cgcacgaatt ggccagcgct gccatttttg
     6181 gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag
     6241 cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac
     6301 agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag
     6361 acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct
     6421 gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca
     6481 agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc
     6541 agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg
     6601 ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc
     6661 tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat
     6721 ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc gcggtgtctc
     6781 gcacacggct tcgacggcgt ttctggcgcg tttgcagggc catagacggc cgccagccca
//
