LOCUS       sequence-349539-        6564 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 5, transcriptional unit for transient 
            expression of iridoid oxidase; CYP76A26 (CrIO) from Catharanthus 
            roseus promoter regions contains 4x attB sites for recruitment fo 
            gal4AD-phiC31.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6564)
  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 6564)
  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: pEPQD1CB0798
FEATURES             Location/Qualifiers
     source          1..6564
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    148..172
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    289..322
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    348..381
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    411..444
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    470..503
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        520..566
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    656..711
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     polyA_signal    2295..2469
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2514..2538
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2694..3842)
                     /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(4178..4766)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4258..4277)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4937..5728,5729..5797))
                     /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     5560..5579
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5798..5902)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5989..6564,1..56)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cagggccata gacggccgcc agcccagcgg cgagggcaac cagcccggtg agcgtcgcaa
       61 aggagatcct gatctgactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg
      121 ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa acaaattgac
      181 gcttagacaa cttaataaca cattgcggac gtttttaatg tactggggtg gatgcagtgg
      241 gccccactct gtgaagacaa cagagaattc gcatgcggag cggtgcgggt gccagggcgt
      301 gcccttgggc tccccgggcg cgtactccac tccccgagcc ggtgcgggtg ccagggcgtg
      361 cccttgggct ccccgggcgc gtactccact ccccccatgg tgcggtgcgg gtgccagggc
      421 gtgcccttgg gctccccggg cgcgtactcc actccccgag ccggtgcggg tgccagggcg
      481 tgcccttggg ctccccgggc gcgtactcca ctccccttcg caagaccctt cctctatata
      541 aggaagttca tttcatttgg agaggactcc ggtattttta caacaattac cacaacaaaa
      601 caaacaacaa acaacattac aatttactat tctagtcgaa tactgtattt ttacaacaat
      661 taccaacaac aacaaacaac aaacaacatt acaattacta tttacaatta caatggcgac
      721 catcactttc gattcactaa acccggtgac cgtcgccatc tcagccggtt ttcttctcct
      781 ccttatcatc tttgtgaaat caagaactgg ttcatccaaa agaaaaccgc cgggtcctcc
      841 aggatggccg attttcggca atatgttcga cctcggagat ttacctcacc aaacgctgta
      901 caagctcaag tccaaatacg ggccgatcgt atggctccag ctcggctcaa ttaatactat
      961 ggttgttcaa aatgcagttt ccgccgccga gctttttaag aaacacgatg tccccttttg
     1021 tgaccgaaaa gtccctgata ctctcactgc cttcaatttc aaccagggct ccctgggtat
     1081 gaatacttac ggcggacatt ggcgagtgct tagacgcctc tgttccatgg agtttttggt
     1141 aaataaacgt atgaacgaaa ctacagatct aagaaggaga attgaggaca atatggtccg
     1201 ttggattgaa gaagattcat tggcatctaa agcacaagga ggaaccggag cggttcaatt
     1261 gagcaggttt ctttttctaa tggcgtttaa tttggtgggt aaccttatgt tatcgagaga
     1321 tttaatggat aataaggatc cagaaggaag ggaatttttt gattgtatga atgagatatt
     1381 ggaattggct ggtactccta atattgctga ttttttgcct ttgttgaaga aattggatcc
     1441 acttggaatg aagaaaagga tggttgataa tatgtctaga actatgaaaa tttcttcaaa
     1501 gtttgttcaa gaaagacttg ataatagaaa agctggaaag attaatgaaa agaaagattt
     1561 cttggatgtt atgcttgaat accaaggtga tggtaaagat ggtcccgata aattcactga
     1621 gcaacatgtc aacattgtca tcatggaaat gttttttgca ggatcagaga caacaagtat
     1681 cagcattgaa tggggattca cagaacttct ccgaaaccca cacgcattca agaaagtaag
     1741 ggaagagatc gacagagtag tgggagtaaa cagaatggtc gaagagagcg acatggagaa
     1801 tttgccgtat ttgcaagcag ttgtaaagga aacgcttaga ttgcatccgg cacttccaat
     1861 gcttctacca agaaacacaa tggaggacac tgaatacatg ggatacctca ttcctaaagg
     1921 cacacaagtt ttcgtgaatg catgggcaat cggaagggat cctgagtact ggcaggaccc
     1981 tttgagtttc aaaccagaaa ggtttatcaa ttccagtgtt gaatataaag gacaacattt
     2041 tgaattgatt ccttttggtt ctggaagaag aatttgtgtg ggatttcctt tggctcatag
     2101 agttgttcat ctcactttgg ctacattagt ccaggctttt gattgggatc ttggtgctgg
     2161 cgttaaacca caagatattg atttggaaga gagattggga ttgactctga ggaagaagaa
     2221 tccattgaat gtcatcccta agaagagagt tcatatctaa gcttctctag ctagagtcga
     2281 tcgacaagct cgagtttctc cataataatg tgtgagtagt tcccagataa gggaattagg
     2341 gttcctatag ggtttcgctc atgtgttgag catataagaa acccttagta tgtatttgta
     2401 tttgtaaaat acttctatca ataaaatttc taattcctaa aaccaaaatc cagtactaaa
     2461 atccagatcg cttgtgttgt cttctgcacg aagtggttta aactatcagt gtttgacagg
     2521 atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa
     2581 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
     2641 ccagatcagg cgctggctgc tgaaccccca gccggaactg accccacaag gccctagcgt
     2701 ttgcaatgca ccaggtcatc attgacccag gcgtgttcca ccaggccgct gcctcgcaac
     2761 tcttcgcagg cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga atccgatccg
     2821 cacatgaggc ggaaggtttc cagcttgagc gggtacggct cccggtgcga gctgaaatag
     2881 tcgaacatcc gtcgggccgt cggcgacagc ttgcggtact tctcccatat gaatttcgtg
     2941 tagtggtcgc cagcaaacag cacgacgatt tcctcgtcga tcaggacctg gcaacgggac
     3001 gttttcttgc cacggtccag gacgcggaag cggtgcagca gcgacaccga ttccaggtgc
     3061 ccaacgcggt cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag gcattcctcg
     3121 gccttcgtgt aataccggcc attgatcgac cagcccaggt cctggcaaag ctcgtagaac
     3181 gtgaaggtga tcggctcgcc gataggggtg cgcttcgcgt actccaacac ctgctgccac
     3241 accagttcgt catcgtcggc ccgcagctcg acgccggtgt aggtgatctt cacgtccttg
     3301 ttgacgtgga aaatgacctt gttttgcagc gcctcgcgcg ggattttctt gttgcgcgtg
     3361 gtgaacaggg cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc cggccacggc
     3421 gcaatatcga acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg tttcagcaac
     3481 gcggcctgct tggcctcgct gacctgtttt gccaggtcct cgccggcggt ttttcgcttc
     3541 ttggtcgtca tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc tgccgcctcc
     3601 tgttcaagac gacgcgaacg ctccacggcg gccgatggcg cgggcagggc agggggagcc
     3661 agttgcacgc tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga gccgacggac
     3721 tggaaggttt cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc ggcatcctcg
     3781 gcggaaaacc ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa cgtccgattc
     3841 attcaccctc cttgcgggat tgccccgact cacgccgggg caatgtgccc ttattcctga
     3901 tttgacccgc ctggtgcctt ggtgtccaga taatccacct tatcggcaat gaagtcggtc
     3961 ccgtagaccg tctggccgtc cttctcgtac ttggtattcc gaatcttgcc ctgcacgaat
     4021 accagcgacc ccttgcccaa atacttgccg tgggcctcgg cctgagagcc aaaacacttg
     4081 atgcggaaga agtcggtgcg ctcctgcttg tcgccggcat cgttgcgcca catctaggat
     4141 ctgccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4201 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4261 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4321 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4381 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4441 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4501 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4561 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4621 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4681 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4741 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4801 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4861 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4921 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4981 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5041 gcttaccatc tggccccagt gctgcaatga taccgcgaga accacgctca ccggctccag
     5101 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5161 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5221 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5281 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5341 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5401 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5461 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5521 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5581 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5641 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5701 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5761 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5821 gaagcattta tcaaggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5881 ataaacaaat aggggttccg cgcacgaatt ggccagcgct gccatttttg gggtgaggcc
     5941 gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag cgggccggga
     6001 gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac agggcgcagc
     6061 cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag acaggttagc
     6121 ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct gtggacagcc
     6181 cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca agtgtcaagg
     6241 atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc agggcactta
     6301 tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg ttttcgccga
     6361 tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc tcatctgtca
     6421 acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga
     6481 gggccaagtt ttccgcgagg tatccacaac gccggcggcc gcggtgtctc gcacacggct
     6541 tcgacggcgt ttctggcgcg tttg
//
