LOCUS       sequence-349525-        6777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 1, transcriptional unit for transient 
            expression of tryptophan decarboxylase (CrTDC) from Catharanthus 
            roseus driven by 35S promoter; contains a chloroplast transit 
            peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6777)
  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 6777)
  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: pEPQD1CB0774
FEATURES             Location/Qualifiers
     source          1..6777
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    211..235
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        421..766
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     709..729
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2571..2745
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2790..2814
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2970..4118)
                     /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(4454..5042)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4534..4553)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5213..6004,6005..6073))
                     /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     5836..5855
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6074..6178)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6265..6777,1..119)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 atccacaacg ccggcggccg cggtgtctcg cacacggctt cgacggcgtt tctggcgcgt
       61 ttgcagggcc atagacggcc gccagcccag cggcgagggc aaccagcccg gtgagcgtcg
      121 caaaggagat cctgatctga ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
      181 ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaatt
      241 gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg gtggatgcag
      301 tgggccccac tctgtgaaga caatgccgaa ttcggatccg gaggtcaaca tggtggagca
      361 cgacactctg gtctactcca aaaatgtcaa agatacagtc tcagaagatc aaagggctat
      421 tgagactttt caacaaagga taatttcggg aaacctcctc ggattccatt gcccagctat
      481 ctgtcacttc atcgaaagga cagtagaaaa ggaaggtggc tcctacaaat gccatcattg
      541 cgataaagga aaggctatca ttcaagatct ctctgccgac agtggtccca aagatggacc
      601 cccacccacg aggagcatcg tggaaaaaga agaggttcca accacgtcta caaagcaagt
      661 ggattgatgt gacatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
      721 agacccttcc tctatataag gaagttcatt tcatttggag aggacacgct actgtataag
      781 agctcatttt tacaacaatt accaacaaca acaaacaaca aacaacatta caattacatt
      841 tacaattatc gataacatgg cttcttctat gctttcttct gctgctgttg ttgctactcg
      901 tgctagtgct gctcaagcta gtatggttgc tccttttact ggacttaagt ctgctgcttc
      961 ttttcctgtt actagaaagc aaaacaacct tgatattact tctattgcta gtaacggagg
     1021 aagagttcga gcaatgggca gcattgattc aacaaatgta gccatgtcca attctccagt
     1081 tggagaattt aagccacttg aagctgagga attccgaaaa caagcccatc gtatggtaga
     1141 tttcatagcc gattattaca aaaatgtgga aacatatccg gtccttagcg aagtcgaacc
     1201 tggatatctc cgaaaacgta tccccgaaac cgctccttac ctccccgaac cacttgacga
     1261 catcatgaaa gatattcaga aggatattat cccaggaatg acaaattgga tgagccctaa
     1321 tttttatgca ttttttcctg ccactgttag ttcagctgcc tttttaggag aaatgttgtc
     1381 tactgcccta aattcagtag gctttacttg ggtttcttca ccagccgcca ccgaattaga
     1441 aatgattgtt atggattggt tggctcagat ccttaaactc cccaaatctt tcatgttttc
     1501 aggtaccggt ggcggcgtca tccaaaacac cactagcgag tccattcttt gtacaatcat
     1561 tgccgcccgg gaaagggccc tggagaagct cggtcccgat agtattggaa aacttgtctg
     1621 ttacggatcc gatcaaaccc ataccatgtt ccccaaaact tgcaaattgg cgggaattta
     1681 tccgaataat attaggttaa tacctacgac cgtcgaaacg gatttcggca tctcacctca
     1741 agttctacga aaaatggtcg aggatgacgt ggcggccgga tatgtaccgc tgttcttatg
     1801 cgctaccctg ggtaccacct cgaccacggc taccgatcct gtggactcac tttctgaaat
     1861 cgctaacgag tttggtattt ggatccacgt ggatgctgct tatgcgggaa gcgcctgtat
     1921 atgtcccgag tttagacatt acttggatgg aatcgaacga gttgactcac tgagtctgag
     1981 tccacacaaa tggctactcg cttacttaga ttgcacttgc ttgtgggtca agcaaccaca
     2041 tttgttacta agggcactca ctacgaatcc tgagtattta aaaaataaac agagtgattt
     2101 agacaaagtt gtggacttca aaaattggca aatcgcaacg ggacgaaaat ttcggtcgct
     2161 gaaactttgg ctcattttac gtagctatgg agttgttaat ttacagagtc atattcgttc
     2221 tgacgtcgca atgggcaaaa tgttcgaaga atgggttaga tcagactcca gattcgaaat
     2281 tgtggtaccg agaaactttt ctcttgtttg ttttagatta aaacctgacg tttcgagttt
     2341 acatgtagaa gaagtgaata agaaactttt ggacatgctt aactcgacgg gacgagttta
     2401 tatgactcat actattgtgg gaggcatata catgctaaga ctggctgttg gctcgtcact
     2461 aactgaagaa catcatgtac gccgtgtttg ggatttgatt caaaaattaa ccgatgattt
     2521 gctcaaagaa gcttgagctt ctctagctag agtcgatcga caagctcgag tttctccata
     2581 ataatgtgtg agtagttccc agataaggga attagggttc ctatagggtt tcgctcatgt
     2641 gttgagcata taagaaaccc ttagtatgta tttgtatttg taaaatactt ctatcaataa
     2701 aatttctaat tcctaaaacc aaaatccagt actaaaatcc agatcgctgc aattgtcttc
     2761 tgcacgaagt ggtttaaact atcagtgttt gacaggatat attggcgggt aaacctaaga
     2821 gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg tttatccgtt
     2881 cgtccatttg tatgtgcatg ccaaccacag ggttccccag atcaggcgct ggctgctgaa
     2941 cccccagccg gaactgaccc cacaaggccc tagcgtttgc aatgcaccag gtcatcattg
     3001 acccaggcgt gttccaccag gccgctgcct cgcaactctt cgcaggcttc gccgacctgc
     3061 tcgcgccact tcttcacgcg ggtggaatcc gatccgcaca tgaggcggaa ggtttccagc
     3121 ttgagcgggt acggctcccg gtgcgagctg aaatagtcga acatccgtcg ggccgtcggc
     3181 gacagcttgc ggtacttctc ccatatgaat ttcgtgtagt ggtcgccagc aaacagcacg
     3241 acgatttcct cgtcgatcag gacctggcaa cgggacgttt tcttgccacg gtccaggacg
     3301 cggaagcggt gcagcagcga caccgattcc aggtgcccaa cgcggtcgga cgtgaagccc
     3361 atcgccgtcg cctgtaggcg cgacaggcat tcctcggcct tcgtgtaata ccggccattg
     3421 atcgaccagc ccaggtcctg gcaaagctcg tagaacgtga aggtgatcgg ctcgccgata
     3481 ggggtgcgct tcgcgtactc caacacctgc tgccacacca gttcgtcatc gtcggcccgc
     3541 agctcgacgc cggtgtaggt gatcttcacg tccttgttga cgtggaaaat gaccttgttt
     3601 tgcagcgcct cgcgcgggat tttcttgttg cgcgtggtga acagggcaga gcgggccgtg
     3661 tcgtttggca tcgctcgcat cgtgtccggc cacggcgcaa tatcgaacaa ggaaagctgc
     3721 atttccttga tctgctgctt cgtgtgtttc agcaacgcgg cctgcttggc ctcgctgacc
     3781 tgttttgcca ggtcctcgcc ggcggttttt cgcttcttgg tcgtcatagt tcctcgcgtg
     3841 tcgatggtca tcgacttcgc caaacctgcc gcctcctgtt caagacgacg cgaacgctcc
     3901 acggcggccg atggcgcggg cagggcaggg ggagccagtt gcacgctgtc gcgctcgatc
     3961 ttggccgtag cttgctggac catcgagccg acggactgga aggtttcgcg gggcgcacgc
     4021 atgacggtgc ggcttgcgat ggtttcggca tcctcggcgg aaaaccccgc gtcgatcagt
     4081 tcttgcctgt atgccttccg gtcaaacgtc cgattcattc accctccttg cgggattgcc
     4141 ccgactcacg ccggggcaat gtgcccttat tcctgatttg acccgcctgg tgccttggtg
     4201 tccagataat ccaccttatc ggcaatgaag tcggtcccgt agaccgtctg gccgtccttc
     4261 tcgtacttgg tattccgaat cttgccctgc acgaatacca gcgacccctt gcccaaatac
     4321 ttgccgtggg cctcggcctg agagccaaaa cacttgatgc ggaagaagtc ggtgcgctcc
     4381 tgcttgtcgc cggcatcgtt gcgccacatc taggatctgc caggaaccgt aaaaaggccg
     4441 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4501 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4561 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4621 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4681 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4741 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4801 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4861 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4921 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4981 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5041 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5101 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5161 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5221 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5281 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5341 caatgatacc gcgagaacca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5401 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5461 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5521 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5581 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5641 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5701 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5761 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5821 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5881 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5941 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6001 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6061 gttgaatact catactcttc ctttttcaat attattgaag catttatcaa ggttattgtc
     6121 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6181 cgaattggcc agcgctgcca tttttggggt gaggccgttc gcggccgagg ggcgcagccc
     6241 ctggggggat gggaggcccg cgttagcggg ccgggagggt tcgagaaggg ggggcacccc
     6301 ccttcggcgt gcgcggtcac gcgcacaggg cgcagccctg gttaaaaaca aggtttataa
     6361 atattggttt aaaagcaggt taaaagacag gttagcggtg gccgaaaaac gggcggaaac
     6421 ccttgcaaat gctggatttt ctgcctgtgg acagcccctc aaatgtcaat aggtgcgccc
     6481 ctcatctgtc agcactctgc ccctcaagtg tcaaggatcg cgcccctcat ctgtcagtag
     6541 tcgcgcccct caagtgtcaa taccgcaggg cacttatccc caggcttgtc cacatcatct
     6601 gtgggaaact cgcgtaaaat caggcgtttt cgccgatttg cgaggctggc cagctccacg
     6661 tcgccggccg aaatcgagcc tgcccctcat ctgtcaacgc cgcgccgggt gagtcggccc
     6721 ctcaagtgtc aacgtccgcc cctcatctgt cagtgagggc caagttttcc gcgaggt
//
