LOCUS       sequence-349535-        7008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 3, transcriptional unit for transient 
            expression of geraniol 8-oxidase (CrG8H) 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 7008)
  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 7008)
  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: pEPQD1CB0794
FEATURES             Location/Qualifiers
     source          1..7008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      1530..2237
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    2286..2310
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2466..3614)
                     /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(3950..4538)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4030..4049)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4709..5500,5501..5569))
                     /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     5332..5351
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5570..5674)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5761..6392
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6484..6508
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    6625..6658
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    6684..6717
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    6747..6780
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    6806..6839
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        6856..6902
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    join(6992..7008,1..39)
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
ORIGIN
        1 caaacaacaa acaacattac aattactatt tacaattaca atggattacc ttaccataat
       61 attaacttta ctatttgcct tgactctcta tgaagccttc agctacctat ccagaagaac
      121 caaaaacctt cctccaggac catcgccatt gccgttcatc ggaagcctcc atttattagg
      181 cgaccaacca cacaaatcct tagcaaaact ttccaaaaaa cacggtccaa ttatgagtct
      241 caaattaggc cagatcacta caatcgtcat atcttcatca acaatggcga aagaagttct
      301 tcaaaaacag gatttagcat tttcaagcag atcagttcca aacgcactcc acgctcacaa
      361 tcaattcaaa ttctccgttg tatggcttcc ggtagcctca cgatggagaa gccttcgaaa
      421 agttttgaat tctaatatat tttccggcaa tcggctcgac gctaatcaac atttgagaac
      481 tagaaaagta caggaactaa ttgcgtattg ccggaaaaat agccagagcg gagaagcggt
      541 tgacgtcggc cgagctgctt ttagaacttc gttgaatttg ttgtcgaatt tgattttttc
      601 aaaggatttg acggatcctt attcggattc tgccaaggaa ttcaaggatt tggtttggaa
      661 tataatggtt gaggcgggga aacctaattt ggtcgatttt tttcccctgc ttgaaaaagt
      721 tgatcctcaa ggtatacgac atcgtatgac gattcacttt ggggaagttc ttaagctttt
      781 tggtggactt gttaatgaaa gattggagca aagaagatca aaaggggaaa aaaatgatgt
      841 gttggatgta cttctaacta ccagccaaga aagccctgag gaaatcgata gaactcacat
      901 tgagcgaatg tgcttggacc tgtttgtagc agggacggac acaacatcaa gcacattaga
      961 atgggcaatg tcagaaatgc ttaaaaaccc agacaaaatg aagaaaaccc aagatgaact
     1021 tgcacaagta atcggcagag gaaaaacaat agaagaatcc gatattaacc gcttacctta
     1081 cttaagatgc gttatgaaag aaaccttaag gatacatcca ccagttccct tcttaattcc
     1141 tcgcaaagtg gaacaaagtg ttgaggtttg tggatacaat gtccctaaag gatcacaagt
     1201 tcttgtgaat gcttgggcaa ttggacgtga tgaaactgtt tgggatgatg ctttggcatt
     1261 caaacccgag agatttatgg aatctgaatt ggatatccgt ggaagagatt tcgagctgat
     1321 tccgttcggt gctggccgaa gaatttgccc agggttgcca ttggcactaa ggactgtgcc
     1381 tttgatgctt ggttctttgt tgaactcttt taattggaag cttgaaggtg ggatggctcc
     1441 aaaagatttg gatatggagg agaagtttgg tattacactg cagaaggctc atcctttgcg
     1501 tgctgtacca agcacccttt aagcttgtcc tgctttaatg agatatgcga gaagcctatg
     1561 atcgcatgat atttgctttc aattctgttg tgcacgttgt aaaaaacctg agcatgtgta
     1621 gctcagatcc ttaccgccgg tttcggttca ttctaatgaa tatatcaccc gttactatcg
     1681 tatttttatg aataatattc tccgttcaat ttactgattg taccctacta cttatatgta
     1741 caatattaaa atgaaaacaa tatattgtgc tgaataggtt tatagcgaca tctatgatag
     1801 agcgccacaa taacaaacaa ttgcgtttta ttattacaaa tccaatttta aaaaaagcgg
     1861 cagaaccggt caaacctaaa agactgatta cataaatctt attcaaattt caaaagtgcc
     1921 ccaggggcta gtatctacga cacaccgagc ggcgaactaa taacgctcac tgaagggaac
     1981 tccggttccc cgccggcgcg catgggtgag attccttgaa gttgagtatt ggccgtccgc
     2041 tctaccgaaa gttacgggca ccattcaacc cggtccagca cggcggccgg gtaaccgact
     2101 tgctgccccg agaattatgc agcatttttt tggtgtatgt gggccccaaa tgaagtgcag
     2161 gtcaaacctt gacagtgacg acaaatcgtt gggcgggtcc agggcgaatt ttgcgacaac
     2221 atgtcgaggc tcagcaggac cgctttactt gtcttctgca cgaagtggtt taaactatca
     2281 gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat
     2341 cggatattta aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa
     2401 ccacagggtt ccccagatca ggcgctggct gctgaacccc cagccggaac tgaccccaca
     2461 aggccctagc gtttgcaatg caccaggtca tcattgaccc aggcgtgttc caccaggccg
     2521 ctgcctcgca actcttcgca ggcttcgccg acctgctcgc gccacttctt cacgcgggtg
     2581 gaatccgatc cgcacatgag gcggaaggtt tccagcttga gcgggtacgg ctcccggtgc
     2641 gagctgaaat agtcgaacat ccgtcgggcc gtcggcgaca gcttgcggta cttctcccat
     2701 atgaatttcg tgtagtggtc gccagcaaac agcacgacga tttcctcgtc gatcaggacc
     2761 tggcaacggg acgttttctt gccacggtcc aggacgcgga agcggtgcag cagcgacacc
     2821 gattccaggt gcccaacgcg gtcggacgtg aagcccatcg ccgtcgcctg taggcgcgac
     2881 aggcattcct cggccttcgt gtaataccgg ccattgatcg accagcccag gtcctggcaa
     2941 agctcgtaga acgtgaaggt gatcggctcg ccgatagggg tgcgcttcgc gtactccaac
     3001 acctgctgcc acaccagttc gtcatcgtcg gcccgcagct cgacgccggt gtaggtgatc
     3061 ttcacgtcct tgttgacgtg gaaaatgacc ttgttttgca gcgcctcgcg cgggattttc
     3121 ttgttgcgcg tggtgaacag ggcagagcgg gccgtgtcgt ttggcatcgc tcgcatcgtg
     3181 tccggccacg gcgcaatatc gaacaaggaa agctgcattt ccttgatctg ctgcttcgtg
     3241 tgtttcagca acgcggcctg cttggcctcg ctgacctgtt ttgccaggtc ctcgccggcg
     3301 gtttttcgct tcttggtcgt catagttcct cgcgtgtcga tggtcatcga cttcgccaaa
     3361 cctgccgcct cctgttcaag acgacgcgaa cgctccacgg cggccgatgg cgcgggcagg
     3421 gcagggggag ccagttgcac gctgtcgcgc tcgatcttgg ccgtagcttg ctggaccatc
     3481 gagccgacgg actggaaggt ttcgcggggc gcacgcatga cggtgcggct tgcgatggtt
     3541 tcggcatcct cggcggaaaa ccccgcgtcg atcagttctt gcctgtatgc cttccggtca
     3601 aacgtccgat tcattcaccc tccttgcggg attgccccga ctcacgccgg ggcaatgtgc
     3661 ccttattcct gatttgaccc gcctggtgcc ttggtgtcca gataatccac cttatcggca
     3721 atgaagtcgg tcccgtagac cgtctggccg tccttctcgt acttggtatt ccgaatcttg
     3781 ccctgcacga ataccagcga ccccttgccc aaatacttgc cgtgggcctc ggcctgagag
     3841 ccaaaacact tgatgcggaa gaagtcggtg cgctcctgct tgtcgccggc atcgttgcgc
     3901 cacatctagg atctgccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     3961 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4021 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4081 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4141 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4201 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4261 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4321 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4381 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4441 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4501 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4561 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     4621 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     4681 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     4741 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     4801 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gaaccacgct
     4861 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     4921 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     4981 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5041 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5101 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5161 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5221 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5281 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5341 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5401 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5461 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     5521 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     5581 ttcaatatta ttgaagcatt tatcaaggtt attgtctcat gagcggatac atatttgaat
     5641 gtatttagaa aaataaacaa ataggggttc cgcgcacgaa ttggccagcg ctgccatttt
     5701 tggggtgagg ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt
     5761 agcgggccgg gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc
     5821 acagggcgca gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa
     5881 agacaggtta gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc
     5941 ctgtggacag cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct
     6001 caagtgtcaa ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc
     6061 gcagggcact tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg
     6121 cgttttcgcc gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc
     6181 cctcatctgt caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc
     6241 atctgtcagt gagggccaag ttttccgcga ggtatccaca acgccggcgg ccgcggtgtc
     6301 tcgcacacgg cttcgacggc gtttctggcg cgtttgcagg gccatagacg gccgccagcc
     6361 cagcggcgag ggcaaccagc ccggtgagcg tcgcaaagga gatcctgatc tgactgatgg
     6421 gctgcctgta tcgagtggtg attttgtgcc gagctgccgg tcggggagct gttggctggc
     6481 tggtggcagg atatattgtg gtgtaaacaa attgacgctt agacaactta ataacacatt
     6541 gcggacgttt ttaatgtact ggggtggatg cagtgggccc cactctgtga agacaaacta
     6601 gaattcgagc tcggagcggt gcgggtgcca gggcgtgccc ttgggctccc cgggcgcgta
     6661 ctccactccc cgagccggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac
     6721 tccactcccc ccatggtgcg gtgcgggtgc cagggcgtgc ccttgggctc cccgggcgcg
     6781 tactccactc cccgagccgg tgcgggtgcc agggcgtgcc cttgggctcc ccgggcgcgt
     6841 actccactcc ccttcgcaag acccttcctc tatataagga agttcatttc atttggagag
     6901 gactccggta tttttacaac aattaccaca acaaaacaaa caacaaacaa cattacaatt
     6961 tactattcta gtcgaatact gtatttttac aacaattacc aacaacaa
//
