LOCUS       sequence-349537-        6183 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 1, transcriptional unit for transient 
            expression of iridoid synthase (CrISY) 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 6183)
  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 6183)
  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: pEPQD1CB0790
FEATURES             Location/Qualifiers
     source          1..6183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    171..195
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    312..345
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    371..404
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    434..467
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    493..526
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        543..589
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    679..734
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     polyA_signal    1937..2111
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2156..2180
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2336..3484)
                     /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(3820..4408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3900..3919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4579..5370,5371..5439))
                     /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     5202..5221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5440..5544)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5631..6183,1..79)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cgacggcgtt tctggcgcgt ttgcagggcc atagacggcc gccagcccag cggcgagggc
       61 aaccagcccg gtgagcgtcg caaaggagat cctgatctga ctgatgggct gcctgtatcg
      121 agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata
      181 tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta
      241 atgtactggg gtggatgcag tgggccccac tctgtgaaga caatgccgaa ttcggatccg
      301 gagcggtgcg ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc cactccccga
      361 gccggtgcgg gtgccagggc gtgcccttgg gctccccggg cgcgtactcc actcccccca
      421 tggtgcggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac tccactcccc
      481 gagccggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact ccactcccct
      541 tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac tccggtattt
      601 ttacaacaat taccacaaca aaacaaacaa caaacaacat tacaatttac tattctagtc
      661 gaatactgta tttttacaac aattaccaac aacaacaaac aacaaacaac attacaatta
      721 ctatttacaa ttacaatgag ttggtggtgg aagaggtcca ttggtgctgg aaagaactta
      781 ccaaaccaga acaaagaaaa tggagtttgt aaaagctaca agagtgtggc actagtagta
      841 ggagtcaccg gcattgttgg cagtagttta gccgaagttc taaagctacc agacacccct
      901 ggaggtccct ggaaggtata cggcgtggca aggcgcccat gccctgtatg gctagccaaa
      961 aaaccagttg agtacatcca atgtgatgtc tcagataatc aagaaacaat ttctaaatta
     1021 tctccactta aggacataac tcatatattc tatgtttctt ggattggatc agaagattgc
     1081 caaacaaatg ctacaatgtt caaaaacatc cttaattcag tgattcctaa tgcttcaaat
     1141 ctccaacatg tgtgtctcca aacaggaatc aaacattatt ttggtatttt tgaagaaggt
     1201 tctaaagttg taccacatga ttcgcctttt accgaggatt tacctaggct aaatgtccca
     1261 aatttctacc atgatcttga agatatactg tatgaagaaa ctggcaagaa caatttaaca
     1321 tggtctgtcc atagaccagc tctggttttc gggttttccc catgtagtat gatgaacatt
     1381 gtcagtacat tatgcgttta cgccacaatt tgtaaacatg aaaataaggc cttggtttat
     1441 ccaggtagta aaaattcatg gaattgttat gctgatgctg ttgatgcaga tttagtagcc
     1501 gagcacgaaa tttgggcagc agttgatcct aaggctaaaa atcaagtatt gaattgtaac
     1561 aatggggatg tttttaaatg gaaacatatt tggaagaaat tagcagagga atttgggatt
     1621 gaaatggtgg gatatgttga aggtaaagaa caagttagtt tggctgagtt aatgaaggat
     1681 aaagatcaag tttgggatga aattgttaag aagaacaatt tagtacctac taaattgaaa
     1741 gaaattgctg ccttttggtt tgctgatatt gctttttgtt ctgagaattt gattagtagt
     1801 atgaacaaaa gtaaggaatt agggttcttg ggttttagga attcgatgaa atcgtttgtt
     1861 tcttgtattg ataagatgag ggattatagg tttattcctt aggcttctct agctagagtc
     1921 gatcgacaag ctcgagtttc tccataataa tgtgtgagta gttcccagat aagggaatta
     1981 gggttcctat agggtttcgc tcatgtgttg agcatataag aaacccttag tatgtatttg
     2041 tatttgtaaa atacttctat caataaaatt tctaattcct aaaaccaaaa tccagtacta
     2101 aaatccagat cgctgcaatt gtcttctgca cgaagtggtt taaactatca gtgtttgaca
     2161 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
     2221 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
     2281 ccccagatca ggcgctggct gctgaacccc cagccggaac tgaccccaca aggccctagc
     2341 gtttgcaatg caccaggtca tcattgaccc aggcgtgttc caccaggccg ctgcctcgca
     2401 actcttcgca ggcttcgccg acctgctcgc gccacttctt cacgcgggtg gaatccgatc
     2461 cgcacatgag gcggaaggtt tccagcttga gcgggtacgg ctcccggtgc gagctgaaat
     2521 agtcgaacat ccgtcgggcc gtcggcgaca gcttgcggta cttctcccat atgaatttcg
     2581 tgtagtggtc gccagcaaac agcacgacga tttcctcgtc gatcaggacc tggcaacggg
     2641 acgttttctt gccacggtcc aggacgcgga agcggtgcag cagcgacacc gattccaggt
     2701 gcccaacgcg gtcggacgtg aagcccatcg ccgtcgcctg taggcgcgac aggcattcct
     2761 cggccttcgt gtaataccgg ccattgatcg accagcccag gtcctggcaa agctcgtaga
     2821 acgtgaaggt gatcggctcg ccgatagggg tgcgcttcgc gtactccaac acctgctgcc
     2881 acaccagttc gtcatcgtcg gcccgcagct cgacgccggt gtaggtgatc ttcacgtcct
     2941 tgttgacgtg gaaaatgacc ttgttttgca gcgcctcgcg cgggattttc ttgttgcgcg
     3001 tggtgaacag ggcagagcgg gccgtgtcgt ttggcatcgc tcgcatcgtg tccggccacg
     3061 gcgcaatatc gaacaaggaa agctgcattt ccttgatctg ctgcttcgtg tgtttcagca
     3121 acgcggcctg cttggcctcg ctgacctgtt ttgccaggtc ctcgccggcg gtttttcgct
     3181 tcttggtcgt catagttcct cgcgtgtcga tggtcatcga cttcgccaaa cctgccgcct
     3241 cctgttcaag acgacgcgaa cgctccacgg cggccgatgg cgcgggcagg gcagggggag
     3301 ccagttgcac gctgtcgcgc tcgatcttgg ccgtagcttg ctggaccatc gagccgacgg
     3361 actggaaggt ttcgcggggc gcacgcatga cggtgcggct tgcgatggtt tcggcatcct
     3421 cggcggaaaa ccccgcgtcg atcagttctt gcctgtatgc cttccggtca aacgtccgat
     3481 tcattcaccc tccttgcggg attgccccga ctcacgccgg ggcaatgtgc ccttattcct
     3541 gatttgaccc gcctggtgcc ttggtgtcca gataatccac cttatcggca atgaagtcgg
     3601 tcccgtagac cgtctggccg tccttctcgt acttggtatt ccgaatcttg ccctgcacga
     3661 ataccagcga ccccttgccc aaatacttgc cgtgggcctc ggcctgagag ccaaaacact
     3721 tgatgcggaa gaagtcggtg cgctcctgct tgtcgccggc atcgttgcgc cacatctagg
     3781 atctgccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3841 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3901 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3961 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     4021 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     4081 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     4141 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     4201 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     4261 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     4321 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     4381 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     4441 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     4501 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     4561 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4621 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4681 gggcttacca tctggcccca gtgctgcaat gataccgcga gaaccacgct caccggctcc
     4741 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4801 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4861 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4921 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4981 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     5041 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     5101 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     5161 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     5221 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     5281 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     5341 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     5401 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     5461 ttgaagcatt tatcaaggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     5521 aaataaacaa ataggggttc cgcgcacgaa ttggccagcg ctgccatttt tggggtgagg
     5581 ccgttcgcgg ccgaggggcg cagcccctgg ggggatggga ggcccgcgtt agcgggccgg
     5641 gagggttcga gaaggggggg cacccccctt cggcgtgcgc ggtcacgcgc acagggcgca
     5701 gccctggtta aaaacaaggt ttataaatat tggtttaaaa gcaggttaaa agacaggtta
     5761 gcggtggccg aaaaacgggc ggaaaccctt gcaaatgctg gattttctgc ctgtggacag
     5821 cccctcaaat gtcaataggt gcgcccctca tctgtcagca ctctgcccct caagtgtcaa
     5881 ggatcgcgcc cctcatctgt cagtagtcgc gcccctcaag tgtcaatacc gcagggcact
     5941 tatccccagg cttgtccaca tcatctgtgg gaaactcgcg taaaatcagg cgttttcgcc
     6001 gatttgcgag gctggccagc tccacgtcgc cggccgaaat cgagcctgcc cctcatctgt
     6061 caacgccgcg ccgggtgagt cggcccctca agtgtcaacg tccgcccctc atctgtcagt
     6121 gagggccaag ttttccgcga ggtatccaca acgccggcgg ccgcggtgtc tcgcacacgg
     6181 ctt
//
