LOCUS       sequence-349437-        6165 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 4, transcriptional unit for transient 
            expression of 8-hydroxygeraniol oxidoreductase (CrGOR) 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 6165)
  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 6165)
  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: pEPQD1CB0796
FEATURES             Location/Qualifiers
     source          1..6165
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(321..1112,1113..1181))
                     /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     944..963
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1182..1286)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1373..2004
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    2096..2120
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    2237..2270
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    2296..2329
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    2359..2392
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    2418..2451
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        2468..2514
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    2604..2659
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      3817..4017
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    4063..4087
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4243..5391)
                     /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(join(5727..6165,1..150))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5807..5826)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
       61 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
      121 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      181 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
      241 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
      301 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
      361 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
      421 gagggcttac catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct
      481 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
      541 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
      601 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
      661 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
      721 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
      781 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
      841 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
      901 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
      961 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     1021 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     1081 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     1141 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     1201 tattgaagca tttatcaagg ttattgtctc atgagcggat acatatttga atgtatttag
     1261 aaaaataaac aaataggggt tccgcgcacg aattggccag cgctgccatt tttggggtga
     1321 ggccgttcgc ggccgagggg cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc
     1381 gggagggttc gagaaggggg ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg
     1441 cagccctggt taaaaacaag gtttataaat attggtttaa aagcaggtta aaagacaggt
     1501 tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc tggattttct gcctgtggac
     1561 agcccctcaa atgtcaatag gtgcgcccct catctgtcag cactctgccc ctcaagtgtc
     1621 aaggatcgcg cccctcatct gtcagtagtc gcgcccctca agtgtcaata ccgcagggca
     1681 cttatcccca ggcttgtcca catcatctgt gggaaactcg cgtaaaatca ggcgttttcg
     1741 ccgatttgcg aggctggcca gctccacgtc gccggccgaa atcgagcctg cccctcatct
     1801 gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca
     1861 gtgagggcca agttttccgc gaggtatcca caacgccggc ggccgcggtg tctcgcacac
     1921 ggcttcgacg gcgtttctgg cgcgtttgca gggccataga cggccgccag cccagcggcg
     1981 agggcaacca gcccggtgag cgtcgcaaag gagatcctga tctgactgat gggctgcctg
     2041 tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg gctggtggca
     2101 ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca ttgcggacgt
     2161 ttttaatgta ctggggtgga tgcagtgggc cccactctgt gaagacaatt acgaattccc
     2221 atggggagcg gtgcgggtgc cagggcgtgc ccttgggctc cccgggcgcg tactccactc
     2281 cccgagccgg tgcgggtgcc agggcgtgcc cttgggctcc ccgggcgcgt actccactcc
     2341 ccccatggtg cggtgcgggt gccagggcgt gcccttgggc tccccgggcg cgtactccac
     2401 tccccgagcc ggtgcgggtg ccagggcgtg cccttgggct ccccgggcgc gtactccact
     2461 ccccttcgca agacccttcc tctatataag gaagttcatt tcatttggag aggactccgg
     2521 tatttttaca acaattacca caacaaaaca aacaacaaac aacattacaa tttactattc
     2581 tagtcgaata ctgtattttt acaacaatta ccaacaacaa caaacaacaa acaacattac
     2641 aattactatt tacaattaca atgaccaaga ccaattcccc tgctccttct gtcatcactt
     2701 gcaaagctgc tgtggtatgg aaatcagggg agccaccaaa ggtggaagag atacaagttg
     2761 atccaccaaa ggcctcagaa gttaggatta agatgctttg tgccagtttg tgccacactg
     2821 atttccttgc ctgcaatggc cttcctgttc cattgttccc tcgcattcct ggacacgaag
     2881 gagtcggaat gatcgagagc gttggagaaa atgttacgaa cctaaaagaa ggagacatag
     2941 tgatgccact ttacttggga gaatgtgggg aatgcttgaa ttgcaaatca ggaaggacaa
     3001 atttgtgcca caaatatcct ttaggtttta gtggattatt gcttgatgga acatcaagaa
     3061 tgtcaattgg agagcaaaaa gtatatcacc atttcagctg ttcgacatgg tcagagtata
     3121 tagtgattga agcagcttat gcagtgaagg tagatccaag ggtttctctt ccacatgcta
     3181 gctttctttg ctgtggattc accactggtt ttggtgctac ttggagagat gtcaatgttg
     3241 tcaagggctc tactgtggct gttctaggcc ttggtgctgt tggacttggg gctgtgcaag
     3301 gagctaaatc acaaggagca tcaagaatta taggtttgga tatcaacgac aagaaacgtg
     3361 aaaagggaga agcatttgga atgactgaat ttataaatcc aaaaggttca aacaaatcaa
     3421 tttctgaact aataaatgaa gcaactggtg gattaggact tgactatgtc tacgaatgca
     3481 ctggagttcc agctttactc aatgaagcca ttgaatcctc taaagtggga cttggaactg
     3541 cagtattgat tggggcagga cttgaaacaa gtggagaaat caaattcatt cctcttctgt
     3601 gtggtagaac tgttaaaggt tcaatttatg gtggagtaag acctaaatca gacctcccca
     3661 ctttaattga aaagtgcata aacaaggaaa ttccaatgga tgaactaatg actcatgaag
     3721 tttcattgtc tgagataaac aaaggatttg agtaccttaa gcatcctgat tgtgtcaaag
     3781 ttgttatcaa gttctaagct tgatcccccg tcgacagcta gctatatcat caatttatgt
     3841 attacacata atatcgcact cagtctttca tctacggcaa tgtaccagct gatataatca
     3901 gttattgaaa tatttctgaa tttaaacttg catcaataaa tttatgtttt tgcttggact
     3961 ataatacctg acttgttatt ttatcaataa atatttaaac tatatttctt tcaagatcgc
     4021 tcagattgtc ttctgcacga agtggtttaa actatcagtg tttgacagga tatattggcg
     4081 ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa
     4141 aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc cagatcaggc
     4201 gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac
     4261 caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc
     4321 ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg
     4381 gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg
     4441 tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc
     4501 agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc
     4561 acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc
     4621 ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta
     4681 ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat
     4741 cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc
     4801 atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa
     4861 aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc
     4921 agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa
     4981 caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt
     5041 ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat
     5101 agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcaagacg
     5161 acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct
     5221 gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact ggaaggtttc
     5281 gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc
     5341 cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc
     5401 ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat ttgacccgcc
     5461 tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc cgtagaccgt
     5521 ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata ccagcgaccc
     5581 cttgcccaaa tacttgccgt gggcctcggc ctgagagcca aaacacttga tgcggaagaa
     5641 gtcggtgcgc tcctgcttgt cgccggcatc gttgcgccac atctaggatc tgccaggaac
     5701 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5761 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5821 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5881 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5941 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     6001 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     6061 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     6121 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctaga
//
