LOCUS       sequence-349529-        6222 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 5, transcriptional unit for transient 
            expression of loganic acid O-methyltransferase (MsLAMT) from 
            Mitragyna speciosa driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6222)
  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 6222)
  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: pEPQD1CB0911
FEATURES             Location/Qualifiers
     source          1..6222
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    193..217
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        403..748
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     691..711
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    1998..2172
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2217..2241
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2397..3545)
                     /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(3881..4469)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3961..3980)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4640..5431,5432..5500))
                     /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     5263..5282
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5501..5605)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5692..6222,1..101)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg ccatagacgg
       61 ccgccagccc agcggcgagg gcaaccagcc cggtgagcgt cgcaaaggag atcctgatct
      121 gactgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg
      181 ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa
      241 taacacattg cggacgtttt taatgtactg gggtggatgc agtgggcccc actctgtgaa
      301 gacaacagag aattcgcatg cggaggtcaa catggtggag cacgacactc tggtctactc
      361 caaaaatgtc aaagatacag tctcagaaga tcaaagggct attgagactt ttcaacaaag
      421 gataatttcg ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcgaaag
      481 gacagtagaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggctat
      541 cattcaagat ctctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat
      601 cgtggaaaaa gaagaggttc caaccacgtc tacaaagcaa gtggattgat gtgacatctc
      661 cactgacgta agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata
      721 aggaagttca tttcatttgg agaggacacg ctcgagtata agagctcatt tttacaacaa
      781 ttaccaacaa caacaaacaa caaacaacat tacaattaca tttacaatta tcgatacaat
      841 ggccccaaca ctggacacca tcccaactgt agatgtcaag gaaatgcctg aagaagcgca
      901 tcctatgaaa ggcggtgatg acttcaatag ctattctcag aattcttgct atcagagggg
      961 agttatagat gctgcaaaag tagttatcgt tgaagcagta aatgaaaagc tggatctaaa
     1021 gaataatgct aattttgatc catctaagcc tttccacatt gctgattttg ggtgctcaac
     1081 cggccctaac acatattttg ccgtgcaaaa tattgtagaa gccgtggacc aaaaacacaa
     1141 atcccttcac gaaaaaatcc agccagaatt ccatgttttt ttcaatgatc atgtcaacaa
     1201 tgatttcaac attctcttca gatctctccc acaggacagg gattattttg ctgctggggt
     1261 ccctggatca ttttacacaa gggtttttcc caaggccagt gtacattttg ctcattgctc
     1321 gtatgcactt cactggctgt ccaaggttcc taaggaaatt caggacaaga actctcctgc
     1381 ctttaacaga gggagaattc attacactgg gactgaaaaa catgtcgtga aagcttactt
     1441 ttctcaattc caaaaagaca tgggctcatt tctgaaagct agagctcaag aaattgtggg
     1501 aggagggttg gtggtgattc aaattcccgg acttccaagt ggtgaggtcc tcttctctag
     1561 gaccggtgct ggcatgcttc atgctctttt gggatcttcc ttgatggaac tcgtaaattt
     1621 gggaataata accgaagaaa gtgtggactc cttcaacttg cctcaatacc atccatcaat
     1681 tgaggaattg gaaatggtga tagagatgaa caatagtttt accattgaaa aagttggagc
     1741 attgaatcac cccatgaaga acttaccatt tgacgttcaa atgacttctc tgcaaataag
     1801 agctgtcatg gaggggattc tgtctgatca ttttggagga aaaatcttgg atcaattgtt
     1861 cgaaatttac accacgaaac tgcaagaaaa ttacaacgtt ttcgacaagg aaatccgaag
     1921 ggatgctgat ttgttccttg ttctaaaacg taagagcaat tgagcttctc tagctagagt
     1981 cgatcgacaa gctcgagttt ctccataata atgtgtgagt agttcccaga taagggaatt
     2041 agggttccta tagggtttcg ctcatgtgtt gagcatataa gaaaccctta gtatgtattt
     2101 gtatttgtaa aatacttcta tcaataaaat ttctaattcc taaaaccaaa atccagtact
     2161 aaaatccaga tcgcttgtgt tgtcttctgc acgaagtggt ttaaactatc agtgtttgac
     2221 aggatatatt ggcgggtaaa cctaagagaa aagagcgttt attagaataa tcggatattt
     2281 aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat gtgcatgcca accacagggt
     2341 tccccagatc aggcgctggc tgctgaaccc ccagccggaa ctgaccccac aaggccctag
     2401 cgtttgcaat gcaccaggtc atcattgacc caggcgtgtt ccaccaggcc gctgcctcgc
     2461 aactcttcgc aggcttcgcc gacctgctcg cgccacttct tcacgcgggt ggaatccgat
     2521 ccgcacatga ggcggaaggt ttccagcttg agcgggtacg gctcccggtg cgagctgaaa
     2581 tagtcgaaca tccgtcgggc cgtcggcgac agcttgcggt acttctccca tatgaatttc
     2641 gtgtagtggt cgccagcaaa cagcacgacg atttcctcgt cgatcaggac ctggcaacgg
     2701 gacgttttct tgccacggtc caggacgcgg aagcggtgca gcagcgacac cgattccagg
     2761 tgcccaacgc ggtcggacgt gaagcccatc gccgtcgcct gtaggcgcga caggcattcc
     2821 tcggccttcg tgtaataccg gccattgatc gaccagccca ggtcctggca aagctcgtag
     2881 aacgtgaagg tgatcggctc gccgataggg gtgcgcttcg cgtactccaa cacctgctgc
     2941 cacaccagtt cgtcatcgtc ggcccgcagc tcgacgccgg tgtaggtgat cttcacgtcc
     3001 ttgttgacgt ggaaaatgac cttgttttgc agcgcctcgc gcgggatttt cttgttgcgc
     3061 gtggtgaaca gggcagagcg ggccgtgtcg tttggcatcg ctcgcatcgt gtccggccac
     3121 ggcgcaatat cgaacaagga aagctgcatt tccttgatct gctgcttcgt gtgtttcagc
     3181 aacgcggcct gcttggcctc gctgacctgt tttgccaggt cctcgccggc ggtttttcgc
     3241 ttcttggtcg tcatagttcc tcgcgtgtcg atggtcatcg acttcgccaa acctgccgcc
     3301 tcctgttcaa gacgacgcga acgctccacg gcggccgatg gcgcgggcag ggcaggggga
     3361 gccagttgca cgctgtcgcg ctcgatcttg gccgtagctt gctggaccat cgagccgacg
     3421 gactggaagg tttcgcgggg cgcacgcatg acggtgcggc ttgcgatggt ttcggcatcc
     3481 tcggcggaaa accccgcgtc gatcagttct tgcctgtatg ccttccggtc aaacgtccga
     3541 ttcattcacc ctccttgcgg gattgccccg actcacgccg gggcaatgtg cccttattcc
     3601 tgatttgacc cgcctggtgc cttggtgtcc agataatcca ccttatcggc aatgaagtcg
     3661 gtcccgtaga ccgtctggcc gtccttctcg tacttggtat tccgaatctt gccctgcacg
     3721 aataccagcg accccttgcc caaatacttg ccgtgggcct cggcctgaga gccaaaacac
     3781 ttgatgcgga agaagtcggt gcgctcctgc ttgtcgccgg catcgttgcg ccacatctag
     3841 gatctgccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     3901 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     3961 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4021 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4081 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4141 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4201 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4261 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     4321 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     4381 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     4441 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     4501 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     4561 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     4621 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct
     4681 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg
     4741 agggcttacc atctggcccc agtgctgcaa tgataccgcg agaaccacgc tcaccggctc
     4801 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa
     4861 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc
     4921 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt
     4981 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc
     5041 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt
     5101 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc
     5161 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt
     5221 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata
     5281 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga
     5341 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag
     5401 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa
     5461 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt
     5521 attgaagcat ttatcaaggt tattgtctca tgagcggata catatttgaa tgtatttaga
     5581 aaaataaaca aataggggtt ccgcgcacga attggccagc gctgccattt ttggggtgag
     5641 gccgttcgcg gccgaggggc gcagcccctg gggggatggg aggcccgcgt tagcgggccg
     5701 ggagggttcg agaagggggg gcacccccct tcggcgtgcg cggtcacgcg cacagggcgc
     5761 agccctggtt aaaaacaagg tttataaata ttggtttaaa agcaggttaa aagacaggtt
     5821 agcggtggcc gaaaaacggg cggaaaccct tgcaaatgct ggattttctg cctgtggaca
     5881 gcccctcaaa tgtcaatagg tgcgcccctc atctgtcagc actctgcccc tcaagtgtca
     5941 aggatcgcgc ccctcatctg tcagtagtcg cgcccctcaa gtgtcaatac cgcagggcac
     6001 ttatccccag gcttgtccac atcatctgtg ggaaactcgc gtaaaatcag gcgttttcgc
     6061 cgatttgcga ggctggccag ctccacgtcg ccggccgaaa tcgagcctgc ccctcatctg
     6121 tcaacgccgc gccgggtgag tcggcccctc aagtgtcaac gtccgcccct catctgtcag
     6181 tgagggccaa gttttccgcg aggtatccac aacgccggcg gc
//
