LOCUS       sequence-349517-        6390 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 5, transcriptional unit for transient 
            expression of loganic acid O-methyltransferase (CrLAMT) from 
            Catharanthus roseus driven by 35S promoter; contains a chloroplast 
            transit peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6390)
  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 6390)
  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: pEPQD1CB0770
FEATURES             Location/Qualifiers
     source          1..6390
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    186..210
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        396..741
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     684..704
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2159..2333
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2378..2402
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2558..3706)
                     /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(4042..4630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4122..4141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4801..5592,5593..5661))
                     /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     5424..5443
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5662..5766)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5853..6390,1..94)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tctcgcacac ggcttcgacg gcgtttctgg cgcgtttgca gggccataga cggccgccag
       61 cccagcggcg agggcaacca gcccggtgag cgtcgcaaag gagatcctga tctgactgat
      121 gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag ctgttggctg
      181 gctggtggca ggatatattg tggtgtaaac aaattgacgc ttagacaact taataacaca
      241 ttgcggacgt ttttaatgta ctggggtgga tgcagtgggc cccactctgt gaagacaaca
      301 gagaattcgc atgcggaggt caacatggtg gagcacgaca ctctggtcta ctccaaaaat
      361 gtcaaagata cagtctcaga agatcaaagg gctattgaga cttttcaaca aaggataatt
      421 tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcga aaggacagta
      481 gaaaaggaag gtggctccta caaatgccat cattgcgata aaggaaaggc tatcattcaa
      541 gatctctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa
      601 aaagaagagg ttccaaccac gtctacaaag caagtggatt gatgtgacat ctccactgac
      661 gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat ataaggaagt
      721 tcatttcatt tggagaggac acgctactgt ataagagctc atttttacaa caattaccaa
      781 caacaacaaa caacaaacaa cattacaatt acatttacaa ttatcgataa catggcttct
      841 tctatgcttt cttctgctgc tgttgttgct actcgtgcta gtgctgctca agctagtatg
      901 gttgctcctt ttactggact taagtctgct gcttcttttc ctgttactag aaagcaaaac
      961 aaccttgata ttacttctat tgctagtaac ggaggaagag ttcgagcaat ggttgccaca
     1021 attgattcca ttgaaatgcc tgcacttcca acagctgtgg aagcccaccc aatgaaaggt
     1081 ggtgatgact cccacagtta ctcccaaaat tcttgctacc agaaaggagt aattgatgca
     1141 gccaaggcag tgattgttga agcagtgaat gaaaaattgg atcttgaaaa taatccaatc
     1201 tttgatccaa tcaaaccttt ccgcattgca gattttggat gttcaacagg accaaatact
     1261 ttccacgcta tgcaaaacat agtagaatca gttgaaacaa aatacaaatc acttcaaaaa
     1321 acccctgaat tccatgtttt cttcaatgat catgttaaca atgatttcaa tgttctcttt
     1381 agatcccttc caccaaacag ggaatttttt gctgctggtg ttcctggatc tttctacact
     1441 agagtttttc ccaaaaatag cattcatttt gctcattgtt cttatgcact tcattggtta
     1501 tctaaagtgc ccaaggaaat tcaagataaa aattctttgg cttataataa gggaagaatt
     1561 cattacactg gtactgaaaa acatgttgtt aaggcttatt ttggtcaatt ccaaagagat
     1621 tttgaagggt ttttaaaagc aagagctcaa gaaattgttg ttggagggtt gatggtgatt
     1681 caaatacctg ggcttcctag tggtgaagtc cttttctcaa ggactggtgc tggtttgctt
     1741 cacttccttt tgggaacttc cttgatggaa ttggttaaca agggaatcat caatgaagaa
     1801 agtgtagatt ccttcaattt gcctcaatat catccctcag ttgaagattt ggaaatggtg
     1861 atagaaatga acgattgttt cacaattgaa agggttggaa ctttacctca tcccatgaag
     1921 aatttaccat ttgatgttca aaggacttct ttacaagtaa gagcaatcat ggaatgcatt
     1981 ctcactgaac attttgggga aaatattttg gatccattat ttgaaatcta caccaaaaat
     2041 ttgcaagaga atttccatgt ttttgataag gaaattagaa aggatgcaga tttgtacctt
     2101 gtcttgaaac gcaagggaaa ttaagcttct ctagctagag tcgatcgaca agctcgagtt
     2161 tctccataat aatgtgtgag tagttcccag ataagggaat tagggttcct atagggtttc
     2221 gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct
     2281 atcaataaaa tttctaattc ctaaaaccaa aatccagtac taaaatccag atcgcttgtg
     2341 ttgtcttctg cacgaagtgg tttaaactat cagtgtttga caggatatat tggcgggtaa
     2401 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
     2461 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttccccagat caggcgctgg
     2521 ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt
     2581 catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc
     2641 cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg
     2701 tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg
     2761 ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa
     2821 acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt
     2881 ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg
     2941 tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc
     3001 ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct
     3061 cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt
     3121 cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga
     3181 ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc
     3241 gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg
     3301 aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct
     3361 cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc
     3421 ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttca agacgacgcg
     3481 aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc
     3541 gctcgatctt ggccgtagct tgctggacca tcgagccgac ggactggaag gtttcgcggg
     3601 gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt
     3661 cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg
     3721 ggattgcccc gactcacgcc ggggcaatgt gcccttattc ctgatttgac ccgcctggtg
     3781 ccttggtgtc cagataatcc accttatcgg caatgaagtc ggtcccgtag accgtctggc
     3841 cgtccttctc gtacttggta ttccgaatct tgccctgcac gaataccagc gaccccttgc
     3901 ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg
     3961 tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa
     4021 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4081 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4141 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4201 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4261 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4321 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4381 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4441 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4501 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4561 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4621 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4681 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4741 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4801 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4861 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4921 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     4981 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5041 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5101 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5161 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5221 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5281 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5341 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5401 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5461 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5521 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5581 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5641 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaagg
     5701 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5761 tccgcgcacg aattggccag cgctgccatt tttggggtga ggccgttcgc ggccgagggg
     5821 cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg
     5881 ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag
     5941 gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg
     6001 gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag
     6061 gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct
     6121 gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca
     6181 catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca
     6241 gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga
     6301 gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc
     6361 gaggtatcca caacgccggc ggccgcggtg
//
