LOCUS       sequence-349531-        5510 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 1, transcriptional unit for transient 
            expression of P19 suppressor of gene silencing from Tomato Bushy 
            Stunt Virus driven by nos promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5510)
  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 5510)
  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: pEPQD1CB0784
FEATURES             Location/Qualifiers
     source          1..5510
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    272..296
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        481..660
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     misc_feature    696..751
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      1284..1536
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    1584..1608
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1764..2912)
                     /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(3248..3836)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3328..3347)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4007..4798,4799..4867))
                     /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     4630..4649
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4868..4972)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5059..5510,1..180)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg
       61 tatccacaac gccggcggcc gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg
      121 tttgcagggc catagacggc cgccagccca gcggcgaggg caaccagccc ggtgagcgtc
      181 gcaaaggaga tcctgatctg actgatgggc tgcctgtatc gagtggtgat tttgtgccga
      241 gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat
      301 tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactgg ggtggatgca
      361 gtgggcccca ctctgtgaag acaatgccga attcggatcc ggagagcgga gaattaaggg
      421 agtcacgtta tgacccccgc cgatgacgcg ggacaagccg ttttacgttt ggaactgaca
      481 gaaccgcaac gttgaaggag ccactcagcc gcgggtttct ggagtttaat gagctaagca
      541 catacgtcag aaaccattat tgcgcgttca aaagtcgcct aaggtcacta tcagctagca
      601 aatatttctt gtcaaaaatg ctccactgac gttccataaa ttcccctcgg tatccaatta
      661 gagtctcata ttcactctca tactgtattt ttacaacaat taccaacaac aacaaacaac
      721 aaacaacatt acaattacta tttacaatta caatggaacg agctatacaa ggaaacgatg
      781 ctagggaaca agcttatggt gaacgttgga atggaggatc aggaagttcc acttctccct
      841 tcaaacttcc tgacgaaagt ccgagttgga ctgagtggcg gctacataac gatgagacta
      901 tttcgaatca agataatccc cttggtttca aggaaagctg gggtttcggg aaagttgtat
      961 ttaagagata tctcagatac gacgggacgg aaacttcact gcacagagtc cttggatctt
     1021 ggacgggaga ttcggttaac tatgcagcat ctcgatttct cggtttcgac cagatcggat
     1081 gtacctatag tattcggttt cgaggagtta gtgtcaccat ttctggaggg tcgcgaactc
     1141 ttcagcatct cagtgaaatg gcaattcggt ctaagcaaga actgctacag cttaccccag
     1201 tcaaagtgga aagtgatgta tcaagaggat gccctgaagg tgttgaaacc ttcgaagaag
     1261 aaagcgagta agcttgtcaa gcagatcgtt caaacatttg gcaataaagt ttcttaagat
     1321 tgaatcctgt tgccggtctt gcgatgatta tcatataatt tctgttgaat tacgttaagc
     1381 atgtaataat taacatgtaa tgcatgacgt tatttatgag atgggttttt atgattagag
     1441 tcccgcaatt atacatttaa tacgcgatag aaaacaaaat atagcgcgca aactaggata
     1501 aattatcgcg cgcggtgtca tctatgttac tagatcgacg ctgcaattgt cttctgcacg
     1561 aagtggttta aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag
     1621 agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca
     1681 tttgtatgtg catgccaacc acagggttcc ccagatcagg cgctggctgc tgaaccccca
     1741 gccggaactg accccacaag gccctagcgt ttgcaatgca ccaggtcatc attgacccag
     1801 gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg cttcgccgac ctgctcgcgc
     1861 cacttcttca cgcgggtgga atccgatccg cacatgaggc ggaaggtttc cagcttgagc
     1921 gggtacggct cccggtgcga gctgaaatag tcgaacatcc gtcgggccgt cggcgacagc
     1981 ttgcggtact tctcccatat gaatttcgtg tagtggtcgc cagcaaacag cacgacgatt
     2041 tcctcgtcga tcaggacctg gcaacgggac gttttcttgc cacggtccag gacgcggaag
     2101 cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt cggacgtgaa gcccatcgcc
     2161 gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt aataccggcc attgatcgac
     2221 cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga tcggctcgcc gataggggtg
     2281 cgcttcgcgt actccaacac ctgctgccac accagttcgt catcgtcggc ccgcagctcg
     2341 acgccggtgt aggtgatctt cacgtccttg ttgacgtgga aaatgacctt gttttgcagc
     2401 gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg cagagcgggc cgtgtcgttt
     2461 ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga acaaggaaag ctgcatttcc
     2521 ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct tggcctcgct gacctgtttt
     2581 gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca tagttcctcg cgtgtcgatg
     2641 gtcatcgact tcgccaaacc tgccgcctcc tgttcaagac gacgcgaacg ctccacggcg
     2701 gccgatggcg cgggcagggc agggggagcc agttgcacgc tgtcgcgctc gatcttggcc
     2761 gtagcttgct ggaccatcga gccgacggac tggaaggttt cgcggggcgc acgcatgacg
     2821 gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc ccgcgtcgat cagttcttgc
     2881 ctgtatgcct tccggtcaaa cgtccgattc attcaccctc cttgcgggat tgccccgact
     2941 cacgccgggg caatgtgccc ttattcctga tttgacccgc ctggtgcctt ggtgtccaga
     3001 taatccacct tatcggcaat gaagtcggtc ccgtagaccg tctggccgtc cttctcgtac
     3061 ttggtattcc gaatcttgcc ctgcacgaat accagcgacc ccttgcccaa atacttgccg
     3121 tgggcctcgg cctgagagcc aaaacacttg atgcggaaga agtcggtgcg ctcctgcttg
     3181 tcgccggcat cgttgcgcca catctaggat ctgccaggaa ccgtaaaaag gccgcgttgc
     3241 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3301 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3361 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     3421 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     3481 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     3541 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     3601 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     3661 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     3721 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     3781 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     3841 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     3901 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     3961 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4021 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4081 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4141 taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4201 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4261 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4321 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4381 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     4441 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     4501 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     4561 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     4621 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     4681 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     4741 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     4801 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     4861 tactcatact cttccttttt caatattatt gaagcattta tcaaggttat tgtctcatga
     4921 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacgaatt
     4981 ggccagcgct gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg
     5041 ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg
     5101 gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg
     5161 gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc
     5221 aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc
     5281 tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc
     5341 ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga
     5401 aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg
     5461 gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg
//
