LOCUS       sequence-349020-        6711 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 3, transcriptional unit for transient 
            expression of geraniol synthase (CrGES) from Catharanthus roseus 
            driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6711)
  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 6711)
  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: pEPQD1CB0110
FEATURES             Location/Qualifiers
     source          1..6711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    85..109
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        295..640
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     583..603
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             2043..2054
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    2379..2553
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2598..2622
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2778..3926)
                     /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(4262..4850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4342..4361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5021..5812,5813..5881))
                     /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     5644..5663
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5882..5986)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6073..6704
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 agatcctgat ctgactgatg ggctgcctgt atcgagtggt gattttgtgc cgagctgccg
       61 gtcggggagc tgttggctgg ctggtggcag gatatattgt ggtgtaaaca aattgacgct
      121 tagacaactt aataacacat tgcggacgtt tttaatgtac tggggtggat gcagtgggcc
      181 ccactctgtg aagacaaact agaattcgag ctcggaggtc aacatggtgg agcacgacac
      241 tctggtctac tccaaaaatg tcaaagatac agtctcagaa gatcaaaggg ctattgagac
      301 ttttcaacaa aggataattt cgggaaacct cctcggattc cattgcccag ctatctgtca
      361 cttcatcgaa aggacagtag aaaaggaagg tggctcctac aaatgccatc attgcgataa
      421 aggaaaggct atcattcaag atctctctgc cgacagtggt cccaaagatg gacccccacc
      481 cacgaggagc atcgtggaaa aagaagaggt tccaaccacg tctacaaagc aagtggattg
      541 atgtgacatc tccactgacg taagggatga cgcacaatcc cactatcctt cgcaagaccc
      601 ttcctctata taaggaagtt catttcattt ggagaggaca cgctcgagta taagagctca
      661 tttttacaac aattaccaac aacaacaaac aacaaacaac attacaatta catttacaat
      721 tatcgataca atgtctctgc ctttggcaac tccattgatc aaagacaatg aatctctcat
      781 caagttcttg cgccaacccc tggtgcttcc tcatgaggtt gatgacagca caaaaaggag
      841 ggaattgttg gaaagaacaa gaaaagaact agaattaaat gcagaaaaac cattggaggc
      901 cttgaagatg atagatataa ttcaaagatt gggattatca tatcattttg aagatgatat
      961 taattcaatt ctcacaggat tttcaaatat tagcagccaa actcatgaag atctcctcac
     1021 tgcttcactt tgttttcgat tgcttcgaca caatgggcat aagatcaatc ctgatatatt
     1081 ccaaaaattc atggacaaca atggaaagtt taaagattca ttaaaggatg acacattagg
     1141 catgttaagc ttatatgaag cttcatattt gggagccaat ggagaagaaa tattgatgga
     1201 agcccaagaa ttcaccaaaa ctcacctgaa aaactcattg ccagccatgg caccatctct
     1261 ttctaagaag gtttctcaag ctttagagca accaagacat agaagaatgt tgaggttaga
     1321 agctagaaga tttattgaag aatatggtgc tgaaaatgac cataatccag accttcttga
     1381 gcttgcaaaa ttggattata acaaagtcca atctctacac caaatggaat tgtctgagat
     1441 aacaaggtgg tggaaacaat tagggcttgt ggataaactc acctttgctc gagatcgacc
     1501 ccttgaatgc tttctttgga cagtgggatt attaccagag cctaagtatt caggttgcag
     1561 aattgagctt gcaaaaacca tagccatttt gcttgtcatt gatgatatct ttgatactca
     1621 tggtacccta gatgagcttc ttctattcac taatgccatt aaaagatggg atcttgaggc
     1681 catggaagat ttaccagaat atatgagaat ttgttacatg gcattgtaca atactactaa
     1741 tgaaatttgc tataaagttc ttaaggaaaa tggttggagt gttcttcctt acctaaaggc
     1801 aacgtggatt gatatgattg aaggattcat ggttgaagca gaatggttca attctgatta
     1861 tgtaccaaac atggaagaat atgtagaaaa tggagttaga acagcaggat catatatggc
     1921 cttagtccat ttgttctttc taatagggca aggtgtcact gaagataatg tgaaattact
     1981 gattaaaccc tatccaaagc tcttttcctc ctcaggaaga atccttcgcc tttgggatga
     2041 tttgggaact gcaaaggagg aacaagaaag aggagacttg gcatcaagca tacaattgtt
     2101 tatgagagag aaagagataa aatcagaaga agaaggaagg aaaggaatat tggaaattat
     2161 agagaattta tggaaagaat tgaatggaga attagtttat agagaagaaa tgcctcttgc
     2221 aataatcaag acagcattca acatggcaag agcttcccaa gttgtgtatc aacatgaaga
     2281 ggacacctat ttttcaagtg tagataatta tgtaaaagct ttgtttttta caccttgttt
     2341 ttaagcttct ctagctagag tcgatcgaca agctcgagtt tctccataat aatgtgtgag
     2401 tagttcccag ataagggaat tagggttcct atagggtttc gctcatgtgt tgagcatata
     2461 agaaaccctt agtatgtatt tgtatttgta aaatacttct atcaataaaa tttctaattc
     2521 ctaaaaccaa aatccagtac taaaatccag atcgctttac ttgtcttctg cacgaagtgg
     2581 tttaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt
     2641 tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta
     2701 tgtgcatgcc aaccacaggg ttccccagat caggcgctgg ctgctgaacc cccagccgga
     2761 actgacccca caaggcccta gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt
     2821 tccaccaggc cgctgcctcg caactcttcg caggcttcgc cgacctgctc gcgccacttc
     2881 ttcacgcggg tggaatccga tccgcacatg aggcggaagg tttccagctt gagcgggtac
     2941 ggctcccggt gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg
     3001 tacttctccc atatgaattt cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg
     3061 tcgatcagga cctggcaacg ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc
     3121 agcagcgaca ccgattccag gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc
     3181 tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc ggccattgat cgaccagccc
     3241 aggtcctggc aaagctcgta gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc
     3301 gcgtactcca acacctgctg ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg
     3361 gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga ccttgttttg cagcgcctcg
     3421 cgcgggattt tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc gtttggcatc
     3481 gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg aaagctgcat ttccttgatc
     3541 tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct cgctgacctg ttttgccagg
     3601 tcctcgccgg cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc
     3661 gacttcgcca aacctgccgc ctcctgttca agacgacgcg aacgctccac ggcggccgat
     3721 ggcgcgggca gggcaggggg agccagttgc acgctgtcgc gctcgatctt ggccgtagct
     3781 tgctggacca tcgagccgac ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg
     3841 cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat
     3901 gccttccggt caaacgtccg attcattcac cctccttgcg ggattgcccc gactcacgcc
     3961 ggggcaatgt gcccttattc ctgatttgac ccgcctggtg ccttggtgtc cagataatcc
     4021 accttatcgg caatgaagtc ggtcccgtag accgtctggc cgtccttctc gtacttggta
     4081 ttccgaatct tgccctgcac gaataccagc gaccccttgc ccaaatactt gccgtgggcc
     4141 tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg tgcgctcctg cttgtcgccg
     4201 gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4261 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4321 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4381 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4441 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4501 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4561 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4621 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4681 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4741 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4801 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     4861 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     4921 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     4981 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5041 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5101 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5161 gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5221 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5281 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5341 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5401 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5461 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5521 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5581 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5641 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5701 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5761 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5821 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5881 tactcttcct ttttcaatat tattgaagca tttatcaagg ttattgtctc atgagcggat
     5941 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcacg aattggccag
     6001 cgctgccatt tttggggtga ggccgttcgc ggccgagggg cgcagcccct ggggggatgg
     6061 gaggcccgcg ttagcgggcc gggagggttc gagaaggggg ggcacccccc ttcggcgtgc
     6121 gcggtcacgc gcacagggcg cagccctggt taaaaacaag gtttataaat attggtttaa
     6181 aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg gcggaaaccc ttgcaaatgc
     6241 tggattttct gcctgtggac agcccctcaa atgtcaatag gtgcgcccct catctgtcag
     6301 cactctgccc ctcaagtgtc aaggatcgcg cccctcatct gtcagtagtc gcgcccctca
     6361 agtgtcaata ccgcagggca cttatcccca ggcttgtcca catcatctgt gggaaactcg
     6421 cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca gctccacgtc gccggccgaa
     6481 atcgagcctg cccctcatct gtcaacgccg cgccgggtga gtcggcccct caagtgtcaa
     6541 cgtccgcccc tcatctgtca gtgagggcca agttttccgc gaggtatcca caacgccggc
     6601 ggccgcggtg tctcgcacac ggcttcgacg gcgtttctgg cgcgtttgca gggccataga
     6661 cggccgccag cccagcggcg agggcaacca gcccggtgag cgtcgcaaag g
//
