LOCUS       sequence-349541-        6642 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 3, transcriptional unit for transient 
            expression of loganic acid O-methyltransferase (CrLAMT) 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 6642)
  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 6642)
  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: pEPQD1CB0806
FEATURES             Location/Qualifiers
     source          1..6642
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    14..47
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    73..106
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    136..169
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    195..228
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     misc_feature    345..369
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     rep_origin      461..1092
                     /label=oriV
                     /note="incP origin of replication"
     promoter        1179..1283
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1284..1352,1353..2144)
                     /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     complement(1502..1521)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2315..2903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2804..2823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             3239..4387
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    4543..4567
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(4616..5323)
                     /label=OCS terminator
                     /note="octopine synthase terminator"
     misc_feature    6448..6503
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     promoter        complement(6593..6639)
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
ORIGIN
        1 ggggagtgga gtacgcgccc ggggagccca agggcacgcc ctggcacccg caccggctcg
       61 gggagtggag tacgcgcccg gggagcccaa gggcacgccc tggcacccgc accgcaccat
      121 ggggggagtg gagtacgcgc ccggggagcc caagggcacg ccctggcacc cgcaccggct
      181 cggggagtgg agtacgcgcc cggggagccc aagggcacgc cctggcaccc gcaccgctcc
      241 gagctcgaat tctagtttgt cttcacagag tggggcccac tgcatccacc ccagtacatt
      301 aaaaacgtcc gcaatgtgtt attaagttgt ctaagcgtca atttgtttac accacaatat
      361 atcctgccac cagccagcca acagctcccc gaccggcagc tcggcacaaa atcaccactc
      421 gatacaggca gcccatcagt cagatcagga tctcctttgc gacgctcacc gggctggttg
      481 ccctcgccgc tgggctggcg gccgtctatg gccctgcaaa cgcgccagaa acgccgtcga
      541 agccgtgtgc gagacaccgc ggccgccggc gttgtggata cctcgcggaa aacttggccc
      601 tcactgacag atgaggggcg gacgttgaca cttgaggggc cgactcaccc ggcgcggcgt
      661 tgacagatga ggggcaggct cgatttcggc cggcgacgtg gagctggcca gcctcgcaaa
      721 tcggcgaaaa cgcctgattt tacgcgagtt tcccacagat gatgtggaca agcctgggga
      781 taagtgccct gcggtattga cacttgaggg gcgcgactac tgacagatga ggggcgcgat
      841 ccttgacact tgaggggcag agtgctgaca gatgaggggc gcacctattg acatttgagg
      901 ggctgtccac aggcagaaaa tccagcattt gcaagggttt ccgcccgttt ttcggccacc
      961 gctaacctgt cttttaacct gcttttaaac caatatttat aaaccttgtt tttaaccagg
     1021 gctgcgccct gtgcgcgtga ccgcgcacgc cgaagggggg tgccccccct tctcgaaccc
     1081 tcccggcccg ctaacgcggg cctcccatcc ccccaggggc tgcgcccctc ggccgcgaac
     1141 ggcctcaccc caaaaatggc agcgctggcc aattcgtgcg cggaacccct atttgtttat
     1201 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccttga taaatgcttc
     1261 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     1321 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     1381 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     1441 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     1501 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     1561 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     1621 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     1681 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     1741 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     1801 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     1861 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     1921 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     1981 ctggagccgg tgagcgtggt tctcgcggta tcattgcagc actggggcca gatggtaagc
     2041 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     2101 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     2161 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     2221 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2281 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2341 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2401 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2461 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2521 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2581 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2641 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2701 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2761 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2821 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2881 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcag
     2941 atcctagatg tggcgcaacg atgccggcga caagcaggag cgcaccgact tcttccgcat
     3001 caagtgtttt ggctctcagg ccgaggccca cggcaagtat ttgggcaagg ggtcgctggt
     3061 attcgtgcag ggcaagattc ggaataccaa gtacgagaag gacggccaga cggtctacgg
     3121 gaccgacttc attgccgata aggtggatta tctggacacc aaggcaccag gcgggtcaaa
     3181 tcaggaataa gggcacattg ccccggcgtg agtcggggca atcccgcaag gagggtgaat
     3241 gaatcggacg tttgaccgga aggcatacag gcaagaactg atcgacgcgg ggttttccgc
     3301 cgaggatgcc gaaaccatcg caagccgcac cgtcatgcgt gcgccccgcg aaaccttcca
     3361 gtccgtcggc tcgatggtcc agcaagctac ggccaagatc gagcgcgaca gcgtgcaact
     3421 ggctccccct gccctgcccg cgccatcggc cgccgtggag cgttcgcgtc gtcttgaaca
     3481 ggaggcggca ggtttggcga agtcgatgac catcgacacg cgaggaacta tgacgaccaa
     3541 gaagcgaaaa accgccggcg aggacctggc aaaacaggtc agcgaggcca agcaggccgc
     3601 gttgctgaaa cacacgaagc agcagatcaa ggaaatgcag ctttccttgt tcgatattgc
     3661 gccgtggccg gacacgatgc gagcgatgcc aaacgacacg gcccgctctg ccctgttcac
     3721 cacgcgcaac aagaaaatcc cgcgcgaggc gctgcaaaac aaggtcattt tccacgtcaa
     3781 caaggacgtg aagatcacct acaccggcgt cgagctgcgg gccgacgatg acgaactggt
     3841 gtggcagcag gtgttggagt acgcgaagcg cacccctatc ggcgagccga tcaccttcac
     3901 gttctacgag ctttgccagg acctgggctg gtcgatcaat ggccggtatt acacgaaggc
     3961 cgaggaatgc ctgtcgcgcc tacaggcgac ggcgatgggc ttcacgtccg accgcgttgg
     4021 gcacctggaa tcggtgtcgc tgctgcaccg cttccgcgtc ctggaccgtg gcaagaaaac
     4081 gtcccgttgc caggtcctga tcgacgagga aatcgtcgtg ctgtttgctg gcgaccacta
     4141 cacgaaattc atatgggaga agtaccgcaa gctgtcgccg acggcccgac ggatgttcga
     4201 ctatttcagc tcgcaccggg agccgtaccc gctcaagctg gaaaccttcc gcctcatgtg
     4261 cggatcggat tccacccgcg tgaagaagtg gcgcgagcag gtcggcgaag cctgcgaaga
     4321 gttgcgaggc agcggcctgg tggaacacgc ctgggtcaat gatgacctgg tgcattgcaa
     4381 acgctagggc cttgtggggt cagttccggc tgggggttca gcagccagcg cctgatctgg
     4441 ggaaccctgt ggttggcatg cacatacaaa tggacgaacg gataaacctt ttcacgccct
     4501 tttaaatatc cgattattct aataaacgct cttttctctt aggtttaccc gccaatatat
     4561 cctgtcaaac actgatagtt taaaccactt cgtgcagaag acaagtaaag cggtcctgct
     4621 gagcctcgac atgttgtcgc aaaattcgcc ctggacccgc ccaacgattt gtcgtcactg
     4681 tcaaggtttg acctgcactt catttggggc ccacatacac caaaaaaatg ctgcataatt
     4741 ctcggggcag caagtcggtt acccggccgc cgtgctggac cgggttgaat ggtgcccgta
     4801 actttcggta gagcggacgg ccaatactca acttcaagga atctcaccca tgcgcgccgg
     4861 cggggaaccg gagttccctt cagtgagcgt tattagttcg ccgctcggtg tgtcgtagat
     4921 actagcccct ggggcacttt tgaaatttga ataagattta tgtaatcagt cttttaggtt
     4981 tgaccggttc tgccgctttt tttaaaattg gatttgtaat aataaaacgc aattgtttgt
     5041 tattgtggcg ctctatcata gatgtcgcta taaacctatt cagcacaata tattgttttc
     5101 attttaatat tgtacatata agtagtaggg tacaatcagt aaattgaacg gagaatatta
     5161 ttcataaaaa tacgatagta acgggtgata tattcattag aatgaaccga aaccggcggt
     5221 aaggatctga gctacacatg ctcaggtttt ttacaacgtg cacaacagaa ttgaaagcaa
     5281 atatcatgcg atcataggct tctcgcatat ctcattaaag caggacaagc ttaatttccc
     5341 ttgcgtttca agacaaggta caaatctgca tcctttctaa tttccttatc aaaaacatgg
     5401 aaattctctt gcaaattttt ggtgtagatt tcaaataatg gatccaaaat attttcccca
     5461 aaatgttcag tgagaatgca ttccatgatt gctcttactt gtaaagaagt cctttgaaca
     5521 tcaaatggta aattcttcat gggatgaggt aaagttccaa ccctttcaat tgtgaaacaa
     5581 tcgttcattt ctatcaccat ttccaaatct tcaactgagg gatgatattg aggcaaattg
     5641 aaggaatcta cactttcttc attgatgatt cccttgttaa ccaattccat caaggaagtt
     5701 cccaaaagga agtgaagcaa accagcacca gtccttgaga aaaggacttc accactagga
     5761 agcccaggta tttgaatcac catcaaccct ccaacaacaa tttcttgagc tcttgctttt
     5821 aaaaaccctt caaaatctct ttggaattga ccaaaataag ccttaacaac atgtttttca
     5881 gtaccagtgt aatgaattct tcccttatta taagccaaag aatttttatc ttgaatttcc
     5941 ttgggcactt tagataacca atgaagtgca taagaacaat gagcaaaatg aatgctattt
     6001 ttgggaaaaa ctctagtgta gaaagatcca ggaacaccag cagcaaaaaa ttccctgttt
     6061 ggtggaaggg atctaaagag aacattgaaa tcattgttaa catgatcatt gaagaaaaca
     6121 tggaattcag gggttttttg aagtgatttg tattttgttt caactgattc tactatgttt
     6181 tgcatagcgt ggaaagtatt tggtcctgtt gaacatccaa aatctgcaat gcggaaaggt
     6241 ttgattggat caaagattgg attattttca agatccaatt tttcattcac tgcttcaaca
     6301 atcactgcct tggctgcatc aattactcct ttctggtagc aagaattttg ggagtaactg
     6361 tgggagtcat caccaccttt cattgggtgg gcttccacag ctgttggaag tgcaggcatt
     6421 tcaatggaat caattgtggc aaccattgta attgtaaata gtaattgtaa tgttgtttgt
     6481 tgtttgttgt tgttggtaat tgttgtaaaa atacagtatt cgactagaat agtaaattgt
     6541 aatgttgttt gttgtttgtt ttgttgtggt aattgttgta aaaataccgg agtcctctcc
     6601 aaatgaaatg aacttcctta tatagaggaa gggtcttgcg aa
//
