LOCUS       sequence-349419-        6234 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 5, transcriptional unit for transient 
            expression of 8-hydroxygeraniol oxidoreductase (CrGOR) from 
            Catharanthus roseus driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6234)
  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 6234)
  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: pEPQD1CB0115
FEATURES             Location/Qualifiers
     source          1..6234
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    177..201
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        387..732
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     675..695
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    1994..2168
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2213..2237
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2393..3541)
                     /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(3877..4465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3957..3976)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4636..5427,5428..5496))
                     /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     5259..5278
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5497..5601)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5688..6234,1..85)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cggcttcgac ggcgtttctg gcgcgtttgc agggccatag acggccgcca gcccagcggc
       61 gagggcaacc agcccggtga gcgtcgcaaa ggagatcctg atctgactga tgggctgcct
      121 gtatcgagtg gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc
      181 aggatatatt gtggtgtaaa caaattgacg cttagacaac ttaataacac attgcggacg
      241 tttttaatgt actggggtgg atgcagtggg ccccactctg tgaagacaac agagaattcg
      301 catgcggagg tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat
      361 acagtctcag aagatcaaag ggctattgag acttttcaac aaaggataat ttcgggaaac
      421 ctcctcggat tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa
      481 ggtggctcct acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatctctct
      541 gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagag
      601 gttccaacca cgtctacaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat
      661 gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat
      721 ttggagagga cacgctcgag tataagagct catttttaca acaattacca acaacaacaa
      781 acaacaaaca acattacaat tacatttaca attatcgata caatgaccaa gaccaattcc
      841 cctgctcctt ctgtcatcac ttgcaaagct gctgtggtat ggaaatcagg ggagccacca
      901 aaggtggaag agatacaagt tgatccacca aaggcctcag aagttaggat taagatgctt
      961 tgtgccagtt tgtgccacac tgatttcctt gcctgcaatg gccttcctgt tccattgttc
     1021 cctcgcattc ctggacacga aggagtcgga atgatcgaga gcgttggaga aaatgttacg
     1081 aacctaaaag aaggagacat agtgatgcca ctttacttgg gagaatgtgg ggaatgcttg
     1141 aattgcaaat caggaaggac aaatttgtgc cacaaatatc ctttaggttt tagtggatta
     1201 ttgcttgatg gaacatcaag aatgtcaatt ggagagcaaa aagtatatca ccatttcagc
     1261 tgttcgacat ggtcagagta tatagtgatt gaagcagctt atgcagtgaa ggtagatcca
     1321 agggtttctc ttccacatgc tagctttctt tgctgtggat tcaccactgg ttttggtgct
     1381 acttggagag atgtcaatgt tgtcaagggc tctactgtgg ctgttctagg ccttggtgct
     1441 gttggacttg gggctgtgca aggagctaaa tcacaaggag catcaagaat tataggtttg
     1501 gatatcaacg acaagaaacg tgaaaaggga gaagcatttg gaatgactga atttataaat
     1561 ccaaaaggtt caaacaaatc aatttctgaa ctaataaatg aagcaactgg tggattagga
     1621 cttgactatg tctacgaatg cactggagtt ccagctttac tcaatgaagc cattgaatcc
     1681 tctaaagtgg gacttggaac tgcagtattg attggggcag gacttgaaac aagtggagaa
     1741 atcaaattca ttcctcttct gtgtggtaga actgttaaag gttcaattta tggtggagta
     1801 agacctaaat cagacctccc cactttaatt gaaaagtgca taaacaagga aattccaatg
     1861 gatgaactaa tgactcatga agtttcattg tctgagataa acaaaggatt tgagtacctt
     1921 aagcatcctg attgtgtcaa agttgttatc aagttctaag cttctctagc tagagtcgat
     1981 cgacaagctc gagtttctcc ataataatgt gtgagtagtt cccagataag ggaattaggg
     2041 ttcctatagg gtttcgctca tgtgttgagc atataagaaa cccttagtat gtatttgtat
     2101 ttgtaaaata cttctatcaa taaaatttct aattcctaaa accaaaatcc agtactaaaa
     2161 tccagatcgc ttgtgttgtc ttctgcacga agtggtttaa actatcagtg tttgacagga
     2221 tatattggcg ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa
     2281 gggcgtgaaa aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc
     2341 cagatcaggc gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt
     2401 tgcaatgcac caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact
     2461 cttcgcaggc ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc
     2521 acatgaggcg gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt
     2581 cgaacatccg tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt
     2641 agtggtcgcc agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg
     2701 ttttcttgcc acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc
     2761 caacgcggtc ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg
     2821 ccttcgtgta ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg
     2881 tgaaggtgat cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca
     2941 ccagttcgtc atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt
     3001 tgacgtggaa aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg
     3061 tgaacagggc agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg
     3121 caatatcgaa caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg
     3181 cggcctgctt ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct
     3241 tggtcgtcat agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct
     3301 gttcaagacg acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca
     3361 gttgcacgct gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact
     3421 ggaaggtttc gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg
     3481 cggaaaaccc cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca
     3541 ttcaccctcc ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat
     3601 ttgacccgcc tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc
     3661 cgtagaccgt ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata
     3721 ccagcgaccc cttgcccaaa tacttgccgt gggcctcggc ctgagagcca aaacacttga
     3781 tgcggaagaa gtcggtgcgc tcctgcttgt cgccggcatc gttgcgccac atctaggatc
     3841 tgccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3901 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3961 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     4021 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4081 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4141 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4201 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4261 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4321 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4381 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4441 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4501 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4561 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4621 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4681 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4741 cttaccatct ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga
     4801 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4861 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4921 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4981 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     5041 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5101 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5161 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5221 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5281 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5341 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5401 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5461 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5521 aagcatttat caaggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5581 taaacaaata ggggttccgc gcacgaattg gccagcgctg ccatttttgg ggtgaggccg
     5641 ttcgcggccg aggggcgcag cccctggggg gatgggaggc ccgcgttagc gggccgggag
     5701 ggttcgagaa gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc
     5761 ctggttaaaa acaaggttta taaatattgg tttaaaagca ggttaaaaga caggttagcg
     5821 gtggccgaaa aacgggcgga aacccttgca aatgctggat tttctgcctg tggacagccc
     5881 ctcaaatgtc aataggtgcg cccctcatct gtcagcactc tgcccctcaa gtgtcaagga
     5941 tcgcgcccct catctgtcag tagtcgcgcc cctcaagtgt caataccgca gggcacttat
     6001 ccccaggctt gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat
     6061 ttgcgaggct ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa
     6121 cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag
     6181 ggccaagttt tccgcgaggt atccacaacg ccggcggccg cggtgtctcg caca
//
