LOCUS       sequence-349577-        6628 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 5, transcriptional unit for transient 
            expression of 7-deoxyloganic acid hydroxylase (Cr7-DLH) 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 6628)
  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 6628)
  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: pEPQD1CB0869
FEATURES             Location/Qualifiers
     source          1..6628
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    100..124
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    241..274
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    300..333
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    363..396
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    422..455
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        472..518
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    608..663
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      2232..2484
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    2530..2554
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2710..3858)
                     /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(4194..4782)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4274..4293)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4953..5744,5745..5813))
                     /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     5576..5595
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5814..5918)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6005..6628,1..8)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tgagcgtcgc aaaggagatc ctgatctgac tgatgggctg cctgtatcga gtggtgattt
       61 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt
      121 aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa tgtactgggg
      181 tggatgcagt gggccccact ctgtgaagac aacagagaat tcgcatgcgg agcggtgcgg
      241 gtgccagggc gtgcccttgg gctccccggg cgcgtactcc actccccgag ccggtgcggg
      301 tgccagggcg tgcccttggg ctccccgggc gcgtactcca ctccccccat ggtgcggtgc
      361 gggtgccagg gcgtgccctt gggctccccg ggcgcgtact ccactccccg agccggtgcg
      421 ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc cactcccctt cgcaagaccc
      481 ttcctctata taaggaagtt catttcattt ggagaggact ccggtatttt tacaacaatt
      541 accacaacaa aacaaacaac aaacaacatt acaatttact attctagtcg aatactgtat
      601 ttttacaaca attaccaaca acaacaaaca acaaacaaca ttacaattac tatttacaat
      661 tacaatggaa ttgaacttca aatcaattat tttcttggtt tttgttagtc taacccttta
      721 ctgggtctat agaatcttgg attgggtatg gtttaagccc aagaagctgg aaaaatgctt
      781 aagagaacaa gggtttaagg gaaatcctta cagattgttc ttgggagatc agtatgatag
      841 tggaaaattg atcagacaag ccttgactaa acctattggt gttgaagaag atgttaagaa
      901 acgaattgta cctcatatac tcaaaactgt tgggactcat ggcaagaagt cttttatgtg
      961 ggttggaaga ataccaaggg tgaatatcac agatccagag ctgattaaag aggtcttaac
     1021 aaaatactat aagttccaga agaatcatca tgatcttgac ccaattacca aattgctttt
     1081 gactggaatt ggaagcttag aaggtgaccc ttgggctaaa cgccgaaaaa tcatcaatgc
     1141 tgcttttcac tttgaaaagt taaagcttat gcttcctgca ttctatttga gttgccgtga
     1201 tatggtgaca aaatgggata ataaggtccc tgaaggagga tcagcagagg tggatgtgtg
     1261 gcatgatatt gaaaccttaa ctggagatgt aatttcaaga acattatttg gaagtaactt
     1321 cgaggaaggg agaagaatat ttgaactcat gaaagaactt actgctctta ctattgatgt
     1381 cattcgctcg gtttatatcc ctggacagag gtttcttcca actaagagga acaataggat
     1441 gagagcaatt gacaaagaag ttagagtaag aataacagaa attatcaata agaaaatgaa
     1501 ggtaatgaaa tcaggagaag cagcaagtgc tgctgatgat tttttgggaa tattgttgga
     1561 atgcaatctg aatgaaataa aggagcaagg aaataacaaa agtgctggaa tgactattgg
     1621 agagattatt ggtgagtgca aactcttcta ttttgctggc caggacacta cctcgacttt
     1681 actcgtgtgg acaatggtcc tcttgtctag attccccgaa tggcaaactc gtgccagaga
     1741 agaagttttt caagtctttg gtaataaaac cccggactat gatggtatca gccatcttaa
     1801 agttataacg atgatcttgt acgaggtttt gaggttatat actccagtgg ctgaactgac
     1861 aaaagtggct catgaagcta cacaactggg aaaatacttc attccagcgg gcgtgcagct
     1921 aatgatgcca caaatcctac ttcatcatga ccctgaaata tggggtgaag atgtaatgga
     1981 gttcaaacca gaaagatttg cagagggagt tctcaaggca actaagagcc aaggctcttt
     2041 ttttccattt agcttgggac caagaatgtg cattggccaa aactttgctc tattggaagc
     2101 caaaatggca atgtctctaa tcttgcgtag attttctttc gagctttctc cctcctatgt
     2161 tcatgctcct ttcactctca tcacaatgca accccaatat ggtgctcatt tgattctgca
     2221 caaactctaa gcttggactc ccatgttggc aaaggcaacc aaacaaacaa tgaatgatcc
     2281 gctcctgcat atggggcggt ttgagtattt caactgccat ttgggctgaa ttgtagacat
     2341 gctcctgtca gaaattccgt gatcttactc aatattcagt aatctcggcc aatatcctaa
     2401 atgtgcgtgg ctttatctgt ctttgtattg tttcatcaat tcatgtaacg tttgcttttc
     2461 ttatgaattt tcaaataaat tatccgcttg tgttgtcttc tgcacgaagt ggtttaaact
     2521 atcagtgttt gacaggatat attggcgggt aaacctaaga gaaaagagcg tttattagaa
     2581 taatcggata tttaaaaggg cgtgaaaagg tttatccgtt cgtccatttg tatgtgcatg
     2641 ccaaccacag ggttccccag atcaggcgct ggctgctgaa cccccagccg gaactgaccc
     2701 cacaaggccc tagcgtttgc aatgcaccag gtcatcattg acccaggcgt gttccaccag
     2761 gccgctgcct cgcaactctt cgcaggcttc gccgacctgc tcgcgccact tcttcacgcg
     2821 ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc ttgagcgggt acggctcccg
     2881 gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc gacagcttgc ggtacttctc
     2941 ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg acgatttcct cgtcgatcag
     3001 gacctggcaa cgggacgttt tcttgccacg gtccaggacg cggaagcggt gcagcagcga
     3061 caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc atcgccgtcg cctgtaggcg
     3121 cgacaggcat tcctcggcct tcgtgtaata ccggccattg atcgaccagc ccaggtcctg
     3181 gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata ggggtgcgct tcgcgtactc
     3241 caacacctgc tgccacacca gttcgtcatc gtcggcccgc agctcgacgc cggtgtaggt
     3301 gatcttcacg tccttgttga cgtggaaaat gaccttgttt tgcagcgcct cgcgcgggat
     3361 tttcttgttg cgcgtggtga acagggcaga gcgggccgtg tcgtttggca tcgctcgcat
     3421 cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc atttccttga tctgctgctt
     3481 cgtgtgtttc agcaacgcgg cctgcttggc ctcgctgacc tgttttgcca ggtcctcgcc
     3541 ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg tcgatggtca tcgacttcgc
     3601 caaacctgcc gcctcctgtt caagacgacg cgaacgctcc acggcggccg atggcgcggg
     3661 cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc ttggccgtag cttgctggac
     3721 catcgagccg acggactgga aggtttcgcg gggcgcacgc atgacggtgc ggcttgcgat
     3781 ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt tcttgcctgt atgccttccg
     3841 gtcaaacgtc cgattcattc accctccttg cgggattgcc ccgactcacg ccggggcaat
     3901 gtgcccttat tcctgatttg acccgcctgg tgccttggtg tccagataat ccaccttatc
     3961 ggcaatgaag tcggtcccgt agaccgtctg gccgtccttc tcgtacttgg tattccgaat
     4021 cttgccctgc acgaatacca gcgacccctt gcccaaatac ttgccgtggg cctcggcctg
     4081 agagccaaaa cacttgatgc ggaagaagtc ggtgcgctcc tgcttgtcgc cggcatcgtt
     4141 gcgccacatc taggatctgc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4201 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4261 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4321 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4381 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4441 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4501 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4561 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4621 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4681 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4741 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4801 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4861 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4921 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4981 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     5041 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca
     5101 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5161 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5221 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5281 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5341 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5401 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5461 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5521 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5581 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5641 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5701 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5761 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5821 ctttttcaat attattgaag catttatcaa ggttattgtc tcatgagcgg atacatattt
     5881 gaatgtattt agaaaaataa acaaataggg gttccgcgca cgaattggcc agcgctgcca
     5941 tttttggggt gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg
     6001 cgttagcggg ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac
     6061 gcgcacaggg cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt
     6121 taaaagacag gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt
     6181 ctgcctgtgg acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc
     6241 ccctcaagtg tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa
     6301 taccgcaggg cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat
     6361 caggcgtttt cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc
     6421 tgcccctcat ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc
     6481 cctcatctgt cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccgcgg
     6541 tgtctcgcac acggcttcga cggcgtttct ggcgcgtttg cagggccata gacggccgcc
     6601 agcccagcgg cgagggcaac cagcccgg
//
