LOCUS       sequence-349009-        5616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 6, transcriptional unit for transient 
            expression of P19 suppressor of gene silencing from Tomato Bushy 
            Stunt Virus driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5616)
  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 5616)
  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: pEPQD1CB0104
FEATURES             Location/Qualifiers
     source          1..5616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    117..141
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        327..672
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     615..635
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    1316..1490
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    1535..1559
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(1715..2863)
                     /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(3199..3787)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3279..3298)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3958..4749,4750..4818))
                     /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     4581..4600
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4819..4923)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5010..5616,1..25)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gagggcaacc agcccggtga gcgtcgcaaa ggagatcctg atctgactga tgggctgcct
       61 gtatcgagtg gtgattttgt gccgagctgc cggtcgggga gctgttggct ggctggtggc
      121 aggatatatt gtggtgtaaa caaattgacg cttagacaac ttaataacac attgcggacg
      181 tttttaatgt actggggtgg atgcagtggg ccccactctg tgaagacaat gtggaattcc
      241 tcgagggagg tcaacatggt ggagcacgac actctggtct actccaaaaa tgtcaaagat
      301 acagtctcag aagatcaaag ggctattgag acttttcaac aaaggataat ttcgggaaac
      361 ctcctcggat tccattgccc agctatctgt cacttcatcg aaaggacagt agaaaaggaa
      421 ggtggctcct acaaatgcca tcattgcgat aaaggaaagg ctatcattca agatctctct
      481 gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagag
      541 gttccaacca cgtctacaaa gcaagtggat tgatgtgaca tctccactga cgtaagggat
      601 gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag ttcatttcat
      661 ttggagagga cacgctcgag tataagagct catttttaca acaattacca acaacaacaa
      721 acaacaaaca acattacaat tacatttaca attatcgata caatggaacg agctatacaa
      781 ggaaacgatg ctagggaaca agcttatggt gaacgttgga atggaggatc aggaagttcc
      841 acttctccct tcaaacttcc tgacgaaagt ccgagttgga ctgagtggcg gctacataac
      901 gatgagacta tttcgaatca agataatccc cttggtttca aggaaagctg gggtttcggg
      961 aaagttgtat ttaagagata tctcagatac gacgggacgg aaacttcact gcacagagtc
     1021 cttggatctt ggacgggaga ttcggttaac tatgcagcat ctcgatttct cggtttcgac
     1081 cagatcggat gtacctatag tattcggttt cgaggagtta gtgtcaccat ttctggaggg
     1141 tcgcgaactc ttcagcatct cagtgaaatg gcaattcggt ctaagcaaga actgctacag
     1201 cttaccccag tcaaagtgga aagtgatgta tcaagaggat gccctgaagg tgttgaaacc
     1261 ttcgaagaag aaagcgagta agcttctcta gctagagtcg atcgacaagc tcgagtttct
     1321 ccataataat gtgtgagtag ttcccagata agggaattag ggttcctata gggtttcgct
     1381 catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa tacttctatc
     1441 aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc gctgagcttg
     1501 tcttctgcac gaagtggttt aaactatcag tgtttgacag gatatattgg cgggtaaacc
     1561 taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat
     1621 ccgttcgtcc atttgtatgt gcatgccaac cacagggttc cccagatcag gcgctggctg
     1681 ctgaaccccc agccggaact gaccccacaa ggccctagcg tttgcaatgc accaggtcat
     1741 cattgaccca ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag gcttcgccga
     1801 cctgctcgcg ccacttcttc acgcgggtgg aatccgatcc gcacatgagg cggaaggttt
     1861 ccagcttgag cgggtacggc tcccggtgcg agctgaaata gtcgaacatc cgtcgggccg
     1921 tcggcgacag cttgcggtac ttctcccata tgaatttcgt gtagtggtcg ccagcaaaca
     1981 gcacgacgat ttcctcgtcg atcaggacct ggcaacggga cgttttcttg ccacggtcca
     2041 ggacgcggaa gcggtgcagc agcgacaccg attccaggtg cccaacgcgg tcggacgtga
     2101 agcccatcgc cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg taataccggc
     2161 cattgatcga ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg atcggctcgc
     2221 cgataggggt gcgcttcgcg tactccaaca cctgctgcca caccagttcg tcatcgtcgg
     2281 cccgcagctc gacgccggtg taggtgatct tcacgtcctt gttgacgtgg aaaatgacct
     2341 tgttttgcag cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg gcagagcggg
     2401 ccgtgtcgtt tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg aacaaggaaa
     2461 gctgcatttc cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc ttggcctcgc
     2521 tgacctgttt tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc atagttcctc
     2581 gcgtgtcgat ggtcatcgac ttcgccaaac ctgccgcctc ctgttcaaga cgacgcgaac
     2641 gctccacggc ggccgatggc gcgggcaggg cagggggagc cagttgcacg ctgtcgcgct
     2701 cgatcttggc cgtagcttgc tggaccatcg agccgacgga ctggaaggtt tcgcggggcg
     2761 cacgcatgac ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac cccgcgtcga
     2821 tcagttcttg cctgtatgcc ttccggtcaa acgtccgatt cattcaccct ccttgcggga
     2881 ttgccccgac tcacgccggg gcaatgtgcc cttattcctg atttgacccg cctggtgcct
     2941 tggtgtccag ataatccacc ttatcggcaa tgaagtcggt cccgtagacc gtctggccgt
     3001 ccttctcgta cttggtattc cgaatcttgc cctgcacgaa taccagcgac cccttgccca
     3061 aatacttgcc gtgggcctcg gcctgagagc caaaacactt gatgcggaag aagtcggtgc
     3121 gctcctgctt gtcgccggca tcgttgcgcc acatctagga tctgccagga accgtaaaaa
     3181 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3241 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3301 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3361 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3421 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3481 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3541 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3601 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3661 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3721 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3781 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3841 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3901 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     3961 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4021 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4081 tgctgcaatg ataccgcgag aaccacgctc accggctcca gatttatcag caataaacca
     4141 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4201 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4261 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4321 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4381 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4441 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4501 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4561 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4621 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4681 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4741 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4801 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcaaggtta
     4861 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4921 gcgcacgaat tggccagcgc tgccattttt ggggtgaggc cgttcgcggc cgaggggcgc
     4981 agcccctggg gggatgggag gcccgcgtta gcgggccggg agggttcgag aagggggggc
     5041 accccccttc ggcgtgcgcg gtcacgcgca cagggcgcag ccctggttaa aaacaaggtt
     5101 tataaatatt ggtttaaaag caggttaaaa gacaggttag cggtggccga aaaacgggcg
     5161 gaaacccttg caaatgctgg attttctgcc tgtggacagc ccctcaaatg tcaataggtg
     5221 cgcccctcat ctgtcagcac tctgcccctc aagtgtcaag gatcgcgccc ctcatctgtc
     5281 agtagtcgcg cccctcaagt gtcaataccg cagggcactt atccccaggc ttgtccacat
     5341 catctgtggg aaactcgcgt aaaatcaggc gttttcgccg atttgcgagg ctggccagct
     5401 ccacgtcgcc ggccgaaatc gagcctgccc ctcatctgtc aacgccgcgc cgggtgagtc
     5461 ggcccctcaa gtgtcaacgt ccgcccctca tctgtcagtg agggccaagt tttccgcgag
     5521 gtatccacaa cgccggcggc cgcggtgtct cgcacacggc ttcgacggcg tttctggcgc
     5581 gtttgcaggg ccatagacgg ccgccagccc agcggc
//
