LOCUS       sequence-349502-        6822 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 2, transcriptional unit for transient 
            expression of iridoid oxidase; CYP76A26 (CrIO) 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 6822)
  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 6822)
  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: pEPQD1CB0122
FEATURES             Location/Qualifiers
     source          1..6822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    100..124
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        310..655
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     598..618
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2505..2679
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2724..2748
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2904..4052)
                     /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(4388..4976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4468..4487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5147..5938,5939..6007))
                     /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     5770..5789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6008..6112)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6199..6822,1..8)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tgagcgtcgc aaaggagatc ctgatctgac tgatgggctg cctgtatcga gtggtgattt
       61 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt
      121 aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg
      181 tggatgcagt gggccccact ctgtgaagac aagcaagaat tcaagcttgg aggtcaacat
      241 ggtggagcac gacactctgg tctactccaa aaatgtcaaa gatacagtct cagaagatca
      301 aagggctatt gagacttttc aacaaaggat aatttcggga aacctcctcg gattccattg
      361 cccagctatc tgtcacttca tcgaaaggac agtagaaaag gaaggtggct cctacaaatg
      421 ccatcattgc gataaaggaa aggctatcat tcaagatctc tctgccgaca gtggtcccaa
      481 agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gaggttccaa ccacgtctac
      541 aaagcaagtg gattgatgtg acatctccac tgacgtaagg gatgacgcac aatcccacta
      601 tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga ggacacgcta
      661 ctgtataaga gctcattttt acaacaatta ccaacaacaa caaacaacaa acaacattac
      721 aattacattt acaattatcg ataacatggc ttcttctatg ctttcttctg ctgctgttgt
      781 tgctactcgt gctagtgctg ctcaagctag tatggttgct ccttttactg gacttaagtc
      841 tgctgcttct tttcctgtta ctagaaagca aaacaacctt gatattactt ctattgctag
      901 taacggagga agagttcgag caatggcgac catcactttc gattcactaa acccggtgac
      961 cgtcgccatc tcagccggtt ttcttctcct ccttatcatc tttgtgaaat caagaactgg
     1021 ttcatccaaa agaaaaccgc cgggtcctcc aggatggccg attttcggca atatgttcga
     1081 cctcggagat ttacctcacc aaacgctgta caagctcaag tccaaatacg ggccgatcgt
     1141 atggctccag ctcggctcaa ttaatactat ggttgttcaa aatgcagttt ccgccgccga
     1201 gctttttaag aaacacgatg tccccttttg tgaccgaaaa gtccctgata ctctcactgc
     1261 cttcaatttc aaccagggct ccctgggtat gaatacttac ggcggacatt ggcgagtgct
     1321 tagacgcctc tgttccatgg agtttttggt aaataaacgt atgaacgaaa ctacagatct
     1381 aagaaggaga attgaggaca atatggtccg ttggattgaa gaagattcat tggcatctaa
     1441 agcacaagga ggaaccggag cggttcaatt gagcaggttt ctttttctaa tggcgtttaa
     1501 tttggtgggt aaccttatgt tatcgagaga tttaatggat aataaggatc cagaaggaag
     1561 ggaatttttt gattgtatga atgagatatt ggaattggct ggtactccta atattgctga
     1621 ttttttgcct ttgttgaaga aattggatcc acttggaatg aagaaaagga tggttgataa
     1681 tatgtctaga actatgaaaa tttcttcaaa gtttgttcaa gaaagacttg ataatagaaa
     1741 agctggaaag attaatgaaa agaaagattt cttggatgtt atgcttgaat accaaggtga
     1801 tggtaaagat ggtcccgata aattcactga gcaacatgtc aacattgtca tcatggaaat
     1861 gttttttgca ggatcagaga caacaagtat cagcattgaa tggggattca cagaacttct
     1921 ccgaaaccca cacgcattca agaaagtaag ggaagagatc gacagagtag tgggagtaaa
     1981 cagaatggtc gaagagagcg acatggagaa tttgccgtat ttgcaagcag ttgtaaagga
     2041 aacgcttaga ttgcatccgg cacttccaat gcttctacca agaaacacaa tggaggacac
     2101 tgaatacatg ggatacctca ttcctaaagg cacacaagtt ttcgtgaatg catgggcaat
     2161 cggaagggat cctgagtact ggcaggaccc tttgagtttc aaaccagaaa ggtttatcaa
     2221 ttccagtgtt gaatataaag gacaacattt tgaattgatt ccttttggtt ctggaagaag
     2281 aatttgtgtg ggatttcctt tggctcatag agttgttcat ctcactttgg ctacattagt
     2341 ccaggctttt gattgggatc ttggtgctgg cgttaaacca caagatattg atttggaaga
     2401 gagattggga ttgactctga ggaagaagaa tccattgaat gtcatcccta agaagagagt
     2461 tcatatctaa gcttctctag ctagagtcga tcgacaagct cgagtttctc cataataatg
     2521 tgtgagtagt tcccagataa gggaattagg gttcctatag ggtttcgctc atgtgttgag
     2581 catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc
     2641 taattcctaa aaccaaaatc cagtactaaa atccagatcg ctactattgt cttctgcacg
     2701 aagtggttta aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag
     2761 agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca
     2821 tttgtatgtg catgccaacc acagggttcc ccagatcagg cgctggctgc tgaaccccca
     2881 gccggaactg accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag
     2941 gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc
     3001 cacttcttca cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc
     3061 gggtacggct cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc
     3121 ttgcggtact tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt
     3181 tcctcgtcga tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag
     3241 cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc
     3301 gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac
     3361 cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg
     3421 cgcttcgcgt actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg
     3481 acgccggtgt aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc
     3541 gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt
     3601 ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc
     3661 ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct gacctgtttt
     3721 gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg
     3781 gtcatcgact tcgccaaacc tgccgcctcc tgttcaagac gacgcgaacg ctccacggcg
     3841 gccgatggcg cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc
     3901 gtagcttgct ggaccatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg
     3961 gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc
     4021 ctgtatgcct tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccgact
     4081 cacgccgggg caatgtgccc ttattcctga tttgacccgc ctggtgcctt ggtgtccaga
     4141 taatccacct tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac
     4201 ttggtattcc gaatcttgcc ctgcacgaat accagcgacc ccttgcccaa atacttgccg
     4261 tgggcctcgg cctgagagcc aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg
     4321 tcgccggcat cgttgcgcca catctaggat ctgccaggaa ccgtaaaaag gccgcgttgc
     4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4801 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5041 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5101 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5161 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5221 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5281 taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5341 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5401 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5461 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5521 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5581 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5641 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5701 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5761 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5821 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5881 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5941 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6001 tactcatact cttccttttt caatattatt gaagcattta tcaaggttat tgtctcatga
     6061 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacgaatt
     6121 ggccagcgct gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg
     6181 ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg
     6241 gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg
     6301 gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc
     6361 aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc
     6421 tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc
     6481 ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga
     6541 aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg
     6601 gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag
     6661 tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac
     6721 gccggcggcc gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg tttgcagggc
     6781 catagacggc cgccagccca gcggcgaggg caaccagccc gg
//
