LOCUS       sequence-349023-        6888 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; contains a chloroplast transit peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6888)
  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 6888)
  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: pEPQD1CB0112
FEATURES             Location/Qualifiers
     source          1..6888
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    311..335
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        521..866
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     809..829
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             2446..2457
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     polyA_signal    2782..2956
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    3001..3025
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3181..4329)
                     /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(4665..5253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4745..4764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5424..6215,6216..6284))
                     /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     6047..6066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6285..6389)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6476..6888,1..219)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg cccctcaagt gtcaacgtcc
       61 gcccctcatc tgtcagtgag ggccaagttt tccgcgaggt atccacaacg ccggcggccg
      121 cggtgtctcg cacacggctt cgacggcgtt tctggcgcgt ttgcagggcc atagacggcc
      181 gccagcccag cggcgagggc aaccagcccg gtgagcgtcg caaaggagat cctgatctga
      241 ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag ctgccggtcg gggagctgtt
      301 ggctggctgg tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata
      361 acacattgcg gacgttttta atgtactggg gtggatgcag tgggccccac tctgtgaaga
      421 caaactagaa ttcgagctcg gaggtcaaca tggtggagca cgacactctg gtctactcca
      481 aaaatgtcaa agatacagtc tcagaagatc aaagggctat tgagactttt caacaaagga
      541 taatttcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc atcgaaagga
      601 cagtagaaaa ggaaggtggc tcctacaaat gccatcattg cgataaagga aaggctatca
      661 ttcaagatct ctctgccgac agtggtccca aagatggacc cccacccacg aggagcatcg
      721 tggaaaaaga agaggttcca accacgtcta caaagcaagt ggattgatgt gacatctcca
      781 ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc tctatataag
      841 gaagttcatt tcatttggag aggacacgct actgtataag agctcatttt tacaacaatt
      901 accaacaaca acaaacaaca aacaacatta caattacatt tacaattatc gataacatgg
      961 cttcttctat gctttcttct gctgctgttg ttgctactcg tgctagtgct gctcaagcta
     1021 gtatggttgc tccttttact ggacttaagt ctgctgcttc ttttcctgtt actagaaagc
     1081 aaaacaacct tgatattact tctattgcta gtaacggagg aagagttcga gcaatgtctc
     1141 tgcctttggc aactccattg atcaaagaca atgaatctct catcaagttc ttgcgccaac
     1201 ccctggtgct tcctcatgag gttgatgaca gcacaaaaag gagggaattg ttggaaagaa
     1261 caagaaaaga actagaatta aatgcagaaa aaccattgga ggccttgaag atgatagata
     1321 taattcaaag attgggatta tcatatcatt ttgaagatga tattaattca attctcacag
     1381 gattttcaaa tattagcagc caaactcatg aagatctcct cactgcttca ctttgttttc
     1441 gattgcttcg acacaatggg cataagatca atcctgatat attccaaaaa ttcatggaca
     1501 acaatggaaa gtttaaagat tcattaaagg atgacacatt aggcatgtta agcttatatg
     1561 aagcttcata tttgggagcc aatggagaag aaatattgat ggaagcccaa gaattcacca
     1621 aaactcacct gaaaaactca ttgccagcca tggcaccatc tctttctaag aaggtttctc
     1681 aagctttaga gcaaccaaga catagaagaa tgttgaggtt agaagctaga agatttattg
     1741 aagaatatgg tgctgaaaat gaccataatc cagaccttct tgagcttgca aaattggatt
     1801 ataacaaagt ccaatctcta caccaaatgg aattgtctga gataacaagg tggtggaaac
     1861 aattagggct tgtggataaa ctcacctttg ctcgagatcg accccttgaa tgctttcttt
     1921 ggacagtggg attattacca gagcctaagt attcaggttg cagaattgag cttgcaaaaa
     1981 ccatagccat tttgcttgtc attgatgata tctttgatac tcatggtacc ctagatgagc
     2041 ttcttctatt cactaatgcc attaaaagat gggatcttga ggccatggaa gatttaccag
     2101 aatatatgag aatttgttac atggcattgt acaatactac taatgaaatt tgctataaag
     2161 ttcttaagga aaatggttgg agtgttcttc cttacctaaa ggcaacgtgg attgatatga
     2221 ttgaaggatt catggttgaa gcagaatggt tcaattctga ttatgtacca aacatggaag
     2281 aatatgtaga aaatggagtt agaacagcag gatcatatat ggccttagtc catttgttct
     2341 ttctaatagg gcaaggtgtc actgaagata atgtgaaatt actgattaaa ccctatccaa
     2401 agctcttttc ctcctcagga agaatccttc gcctttggga tgatttggga actgcaaagg
     2461 aggaacaaga aagaggagac ttggcatcaa gcatacaatt gtttatgaga gagaaagaga
     2521 taaaatcaga agaagaagga aggaaaggaa tattggaaat tatagagaat ttatggaaag
     2581 aattgaatgg agaattagtt tatagagaag aaatgcctct tgcaataatc aagacagcat
     2641 tcaacatggc aagagcttcc caagttgtgt atcaacatga agaggacacc tatttttcaa
     2701 gtgtagataa ttatgtaaaa gctttgtttt ttacaccttg tttttaagct tctctagcta
     2761 gagtcgatcg acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg
     2821 aattagggtt cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt
     2881 atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag
     2941 tactaaaatc cagatcgctt tacttgtctt ctgcacgaag tggtttaaac tatcagtgtt
     3001 tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga ataatcggat
     3061 atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat gccaaccaca
     3121 gggttcccca gatcaggcgc tggctgctga acccccagcc ggaactgacc ccacaaggcc
     3181 ctagcgtttg caatgcacca ggtcatcatt gacccaggcg tgttccacca ggccgctgcc
     3241 tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc gggtggaatc
     3301 cgatccgcac atgaggcgga aggtttccag cttgagcggg tacggctccc ggtgcgagct
     3361 gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg cggtacttct cccatatgaa
     3421 tttcgtgtag tggtcgccag caaacagcac gacgatttcc tcgtcgatca ggacctggca
     3481 acgggacgtt ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg acaccgattc
     3541 caggtgccca acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc gcgacaggca
     3601 ttcctcggcc ttcgtgtaat accggccatt gatcgaccag cccaggtcct ggcaaagctc
     3661 gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact ccaacacctg
     3721 ctgccacacc agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg tgatcttcac
     3781 gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga ttttcttgtt
     3841 gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc atcgctcgca tcgtgtccgg
     3901 ccacggcgca atatcgaaca aggaaagctg catttccttg atctgctgct tcgtgtgttt
     3961 cagcaacgcg gcctgcttgg cctcgctgac ctgttttgcc aggtcctcgc cggcggtttt
     4021 tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg ccaaacctgc
     4081 cgcctcctgt tcaagacgac gcgaacgctc cacggcggcc gatggcgcgg gcagggcagg
     4141 gggagccagt tgcacgctgt cgcgctcgat cttggccgta gcttgctgga ccatcgagcc
     4201 gacggactgg aaggtttcgc ggggcgcacg catgacggtg cggcttgcga tggtttcggc
     4261 atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc ggtcaaacgt
     4321 ccgattcatt caccctcctt gcgggattgc cccgactcac gccggggcaa tgtgccctta
     4381 ttcctgattt gacccgcctg gtgccttggt gtccagataa tccaccttat cggcaatgaa
     4441 gtcggtcccg tagaccgtct ggccgtcctt ctcgtacttg gtattccgaa tcttgccctg
     4501 cacgaatacc agcgacccct tgcccaaata cttgccgtgg gcctcggcct gagagccaaa
     4561 acacttgatg cggaagaagt cggtgcgctc ctgcttgtcg ccggcatcgt tgcgccacat
     4621 ctaggatctg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4861 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5101 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     5341 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     5401 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5461 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5521 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc acgctcaccg
     5581 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5641 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5701 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5761 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5821 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5881 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5941 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6001 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6061 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6301 tattattgaa gcatttatca aggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6361 tagaaaaata aacaaatagg ggttccgcgc acgaattggc cagcgctgcc atttttgggg
     6421 tgaggccgtt cgcggccgag gggcgcagcc cctgggggga tgggaggccc gcgttagcgg
     6481 gccgggaggg ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca cgcgcacagg
     6541 gcgcagccct ggttaaaaac aaggtttata aatattggtt taaaagcagg ttaaaagaca
     6601 ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt tctgcctgtg
     6661 gacagcccct caaatgtcaa taggtgcgcc cctcatctgt cagcactctg cccctcaagt
     6721 gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca ataccgcagg
     6781 gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa tcaggcgttt
     6841 tcgccgattt gcgaggctgg ccagctccac gtcgccggcc gaaatcga
//
