LOCUS       sequence-349501-        6645 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.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6645)
  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 6645)
  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: pEPQD1CB0121
FEATURES             Location/Qualifiers
     source          1..6645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    78..102
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        288..633
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     576..596
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2306..2480
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2525..2549
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2705..3853)
                     /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(4189..4777)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4269..4288)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4948..5739,5740..5808))
                     /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     5571..5590
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5809..5913)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6000..6631
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gatctgactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg
       61 agctgttggc tggctggtgg caggatatat tgtggtgtaa acaaattgac gcttagacaa
      121 cttaataaca cattgcggac gtttttaatg tactggggtg gatgcagtgg gccccactct
      181 gtgaagacaa gcaagaattc aagcttggag gtcaacatgg tggagcacga cactctggtc
      241 tactccaaaa atgtcaaaga tacagtctca gaagatcaaa gggctattga gacttttcaa
      301 caaaggataa tttcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc
      361 gaaaggacag tagaaaagga aggtggctcc tacaaatgcc atcattgcga taaaggaaag
      421 gctatcattc aagatctctc tgccgacagt ggtcccaaag atggaccccc acccacgagg
      481 agcatcgtgg aaaaagaaga ggttccaacc acgtctacaa agcaagtgga ttgatgtgac
      541 atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga cccttcctct
      601 atataaggaa gttcatttca tttggagagg acacgctcga gtataagagc tcatttttac
      661 aacaattacc aacaacaaca aacaacaaac aacattacaa ttacatttac aattatcgat
      721 acaatggcga ccatcacttt cgattcacta aacccggtga ccgtcgccat ctcagccggt
      781 tttcttctcc tccttatcat ctttgtgaaa tcaagaactg gttcatccaa aagaaaaccg
      841 ccgggtcctc caggatggcc gattttcggc aatatgttcg acctcggaga tttacctcac
      901 caaacgctgt acaagctcaa gtccaaatac gggccgatcg tatggctcca gctcggctca
      961 attaatacta tggttgttca aaatgcagtt tccgccgccg agctttttaa gaaacacgat
     1021 gtcccctttt gtgaccgaaa agtccctgat actctcactg ccttcaattt caaccagggc
     1081 tccctgggta tgaatactta cggcggacat tggcgagtgc ttagacgcct ctgttccatg
     1141 gagtttttgg taaataaacg tatgaacgaa actacagatc taagaaggag aattgaggac
     1201 aatatggtcc gttggattga agaagattca ttggcatcta aagcacaagg aggaaccgga
     1261 gcggttcaat tgagcaggtt tctttttcta atggcgttta atttggtggg taaccttatg
     1321 ttatcgagag atttaatgga taataaggat ccagaaggaa gggaattttt tgattgtatg
     1381 aatgagatat tggaattggc tggtactcct aatattgctg attttttgcc tttgttgaag
     1441 aaattggatc cacttggaat gaagaaaagg atggttgata atatgtctag aactatgaaa
     1501 atttcttcaa agtttgttca agaaagactt gataatagaa aagctggaaa gattaatgaa
     1561 aagaaagatt tcttggatgt tatgcttgaa taccaaggtg atggtaaaga tggtcccgat
     1621 aaattcactg agcaacatgt caacattgtc atcatggaaa tgttttttgc aggatcagag
     1681 acaacaagta tcagcattga atggggattc acagaacttc tccgaaaccc acacgcattc
     1741 aagaaagtaa gggaagagat cgacagagta gtgggagtaa acagaatggt cgaagagagc
     1801 gacatggaga atttgccgta tttgcaagca gttgtaaagg aaacgcttag attgcatccg
     1861 gcacttccaa tgcttctacc aagaaacaca atggaggaca ctgaatacat gggatacctc
     1921 attcctaaag gcacacaagt tttcgtgaat gcatgggcaa tcggaaggga tcctgagtac
     1981 tggcaggacc ctttgagttt caaaccagaa aggtttatca attccagtgt tgaatataaa
     2041 ggacaacatt ttgaattgat tccttttggt tctggaagaa gaatttgtgt gggatttcct
     2101 ttggctcata gagttgttca tctcactttg gctacattag tccaggcttt tgattgggat
     2161 cttggtgctg gcgttaaacc acaagatatt gatttggaag agagattggg attgactctg
     2221 aggaagaaga atccattgaa tgtcatccct aagaagagag ttcatatcta agcttctcta
     2281 gctagagtcg atcgacaagc tcgagtttct ccataataat gtgtgagtag ttcccagata
     2341 agggaattag ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt
     2401 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
     2461 ccagtactaa aatccagatc gctactattg tcttctgcac gaagtggttt aaactatcag
     2521 tgtttgacag gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc
     2581 ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac
     2641 cacagggttc cccagatcag gcgctggctg ctgaaccccc agccggaact gaccccacaa
     2701 ggccctagcg tttgcaatgc accaggtcat cattgaccca ggcgtgttcc accaggccgc
     2761 tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc acgcgggtgg
     2821 aatccgatcc gcacatgagg cggaaggttt ccagcttgag cgggtacggc tcccggtgcg
     2881 agctgaaata gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac ttctcccata
     2941 tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg atcaggacct
     3001 ggcaacggga cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc agcgacaccg
     3061 attccaggtg cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt aggcgcgaca
     3121 ggcattcctc ggccttcgtg taataccggc cattgatcga ccagcccagg tcctggcaaa
     3181 gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg tactccaaca
     3241 cctgctgcca caccagttcg tcatcgtcgg cccgcagctc gacgccggtg taggtgatct
     3301 tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc gggattttct
     3361 tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct cgcatcgtgt
     3421 ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc tgcttcgtgt
     3481 gtttcagcaa cgcggcctgc ttggcctcgc tgacctgttt tgccaggtcc tcgccggcgg
     3541 tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac ttcgccaaac
     3601 ctgccgcctc ctgttcaaga cgacgcgaac gctccacggc ggccgatggc gcgggcaggg
     3661 cagggggagc cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc tggaccatcg
     3721 agccgacgga ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt gcgatggttt
     3781 cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc ttccggtcaa
     3841 acgtccgatt cattcaccct ccttgcggga ttgccccgac tcacgccggg gcaatgtgcc
     3901 cttattcctg atttgacccg cctggtgcct tggtgtccag ataatccacc ttatcggcaa
     3961 tgaagtcggt cccgtagacc gtctggccgt ccttctcgta cttggtattc cgaatcttgc
     4021 cctgcacgaa taccagcgac cccttgccca aatacttgcc gtgggcctcg gcctgagagc
     4081 caaaacactt gatgcggaag aagtcggtgc gctcctgctt gtcgccggca tcgttgcgcc
     4141 acatctagga tctgccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4201 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4261 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4321 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4381 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4441 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4501 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4561 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     4621 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4681 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4741 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4801 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     4861 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     4921 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     4981 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5041 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag aaccacgctc
     5101 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5161 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5221 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     5281 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5341 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5401 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5461 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5521 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5581 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5641 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5701 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5761 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     5821 tcaatattat tgaagcattt atcaaggtta ttgtctcatg agcggataca tatttgaatg
     5881 tatttagaaa aataaacaaa taggggttcc gcgcacgaat tggccagcgc tgccattttt
     5941 ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag gcccgcgtta
     6001 gcgggccggg agggttcgag aagggggggc accccccttc ggcgtgcgcg gtcacgcgca
     6061 cagggcgcag ccctggttaa aaacaaggtt tataaatatt ggtttaaaag caggttaaaa
     6121 gacaggttag cggtggccga aaaacgggcg gaaacccttg caaatgctgg attttctgcc
     6181 tgtggacagc ccctcaaatg tcaataggtg cgcccctcat ctgtcagcac tctgcccctc
     6241 aagtgtcaag gatcgcgccc ctcatctgtc agtagtcgcg cccctcaagt gtcaataccg
     6301 cagggcactt atccccaggc ttgtccacat catctgtggg aaactcgcgt aaaatcaggc
     6361 gttttcgccg atttgcgagg ctggccagct ccacgtcgcc ggccgaaatc gagcctgccc
     6421 ctcatctgtc aacgccgcgc cgggtgagtc ggcccctcaa gtgtcaacgt ccgcccctca
     6481 tctgtcagtg agggccaagt tttccgcgag gtatccacaa cgccggcggc cgcggtgtct
     6541 cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg ccatagacgg ccgccagccc
     6601 agcggcgagg gcaaccagcc cggtgagcgt cgcaaaggag atcct
//
