LOCUS       sequence-349515-        6213 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.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6213)
  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 6213)
  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: pEPQD1CB0769
FEATURES             Location/Qualifiers
     source          1..6213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(135..926,927..995))
                     /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     758..777
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(996..1100)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1187..1818
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    1910..1934
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        2120..2465
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2408..2428
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    3706..3880
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3925..3949
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4105..5253)
                     /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(5589..6177)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5669..5688)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
       61 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
      121 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
      181 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
      241 ttaccatctg gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat
      301 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
      361 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
      421 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
      481 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
      541 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
      601 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
      661 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
      721 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
      781 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
      841 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
      901 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
      961 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1021 agcatttatc aaggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1081 aaacaaatag gggttccgcg cacgaattgg ccagcgctgc catttttggg gtgaggccgt
     1141 tcgcggccga ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg ggccgggagg
     1201 gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc
     1261 tggttaaaaa caaggtttat aaatattggt ttaaaagcag gttaaaagac aggttagcgg
     1321 tggccgaaaa acgggcggaa acccttgcaa atgctggatt ttctgcctgt ggacagcccc
     1381 tcaaatgtca ataggtgcgc ccctcatctg tcagcactct gcccctcaag tgtcaaggat
     1441 cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc
     1501 cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt
     1561 tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac
     1621 gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg
     1681 gccaagtttt ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc acacggcttc
     1741 gacggcgttt ctggcgcgtt tgcagggcca tagacggccg ccagcccagc ggcgagggca
     1801 accagcccgg tgagcgtcgc aaaggagatc ctgatctgac tgatgggctg cctgtatcga
     1861 gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat
     1921 attgtggtgt aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa
     1981 tgtactgggg tggatgcagt gggccccact ctgtgaagac aacagagaat tcgcatgcgg
     2041 aggtcaacat ggtggagcac gacactctgg tctactccaa aaatgtcaaa gatacagtct
     2101 cagaagatca aagggctatt gagacttttc aacaaaggat aatttcggga aacctcctcg
     2161 gattccattg cccagctatc tgtcacttca tcgaaaggac agtagaaaag gaaggtggct
     2221 cctacaaatg ccatcattgc gataaaggaa aggctatcat tcaagatctc tctgccgaca
     2281 gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gaggttccaa
     2341 ccacgtctac aaagcaagtg gattgatgtg acatctccac tgacgtaagg gatgacgcac
     2401 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
     2461 ggacacgctc gagtataaga gctcattttt acaacaatta ccaacaacaa caaacaacaa
     2521 acaacattac aattacattt acaattatcg atacaatggt tgccacaatt gattccattg
     2581 aaatgcctgc acttccaaca gctgtggaag cccacccaat gaaaggtggt gatgactccc
     2641 acagttactc ccaaaattct tgctaccaga aaggagtaat tgatgcagcc aaggcagtga
     2701 ttgttgaagc agtgaatgaa aaattggatc ttgaaaataa tccaatcttt gatccaatca
     2761 aacctttccg cattgcagat tttggatgtt caacaggacc aaatactttc cacgctatgc
     2821 aaaacatagt agaatcagtt gaaacaaaat acaaatcact tcaaaaaacc cctgaattcc
     2881 atgttttctt caatgatcat gttaacaatg atttcaatgt tctctttaga tcccttccac
     2941 caaacaggga attttttgct gctggtgttc ctggatcttt ctacactaga gtttttccca
     3001 aaaatagcat tcattttgct cattgttctt atgcacttca ttggttatct aaagtgccca
     3061 aggaaattca agataaaaat tctttggctt ataataaggg aagaattcat tacactggta
     3121 ctgaaaaaca tgttgttaag gcttattttg gtcaattcca aagagatttt gaagggtttt
     3181 taaaagcaag agctcaagaa attgttgttg gagggttgat ggtgattcaa atacctgggc
     3241 ttcctagtgg tgaagtcctt ttctcaagga ctggtgctgg tttgcttcac ttccttttgg
     3301 gaacttcctt gatggaattg gttaacaagg gaatcatcaa tgaagaaagt gtagattcct
     3361 tcaatttgcc tcaatatcat ccctcagttg aagatttgga aatggtgata gaaatgaacg
     3421 attgtttcac aattgaaagg gttggaactt tacctcatcc catgaagaat ttaccatttg
     3481 atgttcaaag gacttcttta caagtaagag caatcatgga atgcattctc actgaacatt
     3541 ttggggaaaa tattttggat ccattatttg aaatctacac caaaaatttg caagagaatt
     3601 tccatgtttt tgataaggaa attagaaagg atgcagattt gtaccttgtc ttgaaacgca
     3661 agggaaatta agcttctcta gctagagtcg atcgacaagc tcgagtttct ccataataat
     3721 gtgtgagtag ttcccagata agggaattag ggttcctata gggtttcgct catgtgttga
     3781 gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc aataaaattt
     3841 ctaattccta aaaccaaaat ccagtactaa aatccagatc gcttgtgttg tcttctgcac
     3901 gaagtggttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     3961 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
     4021 atttgtatgt gcatgccaac cacagggttc cccagatcag gcgctggctg ctgaaccccc
     4081 agccggaact gaccccacaa ggccctagcg tttgcaatgc accaggtcat cattgaccca
     4141 ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga cctgctcgcg
     4201 ccacttcttc acgcgggtgg aatccgatcc gcacatgagg cggaaggttt ccagcttgag
     4261 cgggtacggc tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg tcggcgacag
     4321 cttgcggtac ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca gcacgacgat
     4381 ttcctcgtcg atcaggacct ggcaacggga cgttttcttg ccacggtcca ggacgcggaa
     4441 gcggtgcagc agcgacaccg attccaggtg cccaacgcgg tcggacgtga agcccatcgc
     4501 cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg taataccggc cattgatcga
     4561 ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc cgataggggt
     4621 gcgcttcgcg tactccaaca cctgctgcca caccagttcg tcatcgtcgg cccgcagctc
     4681 gacgccggtg taggtgatct tcacgtcctt gttgacgtgg aaaatgacct tgttttgcag
     4741 cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg gcagagcggg ccgtgtcgtt
     4801 tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa gctgcatttc
     4861 cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc tgacctgttt
     4921 tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc atagttcctc gcgtgtcgat
     4981 ggtcatcgac ttcgccaaac ctgccgcctc ctgttcaaga cgacgcgaac gctccacggc
     5041 ggccgatggc gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct cgatcttggc
     5101 cgtagcttgc tggaccatcg agccgacgga ctggaaggtt tcgcggggcg cacgcatgac
     5161 ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga tcagttcttg
     5221 cctgtatgcc ttccggtcaa acgtccgatt cattcaccct ccttgcggga ttgccccgac
     5281 tcacgccggg gcaatgtgcc cttattcctg atttgacccg cctggtgcct tggtgtccag
     5341 ataatccacc ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt ccttctcgta
     5401 cttggtattc cgaatcttgc cctgcacgaa taccagcgac cccttgccca aatacttgcc
     5461 gtgggcctcg gcctgagagc caaaacactt gatgcggaag aagtcggtgc gctcctgctt
     5521 gtcgccggca tcgttgcgcc acatctagga tctgccagga accgtaaaaa ggccgcgttg
     5581 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5641 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5701 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5761 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5821 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5881 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5941 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     6001 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     6061 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     6121 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     6181 gatcctttga tcttttctac ggggtctgac gct
//
