LOCUS       sequence-349510-        6723 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 3, transcriptional unit for transient 
            expression of 7-deoxyloganetic acid glucosyl transferase (Cr7-DLGT) 
            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 6723)
  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 6723)
  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: pEPQD1CB0124
FEATURES             Location/Qualifiers
     source          1..6723
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    268..292
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        478..823
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     766..786
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2574..2748
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2793..2817
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2973..4121)
                     /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(4457..5045)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4537..4556)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5216..6007,6008..6076))
                     /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     5839..5858
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6077..6181)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6268..6723,1..176)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 ctcaagtgtc aacgtccgcc cctcatctgt cagtgagggc caagttttcc gcgaggtatc
       61 cacaacgccg gcggccgcgg tgtctcgcac acggcttcga cggcgtttct ggcgcgtttg
      121 cagggccata gacggccgcc agcccagcgg cgagggcaac cagcccggtg agcgtcgcaa
      181 aggagatcct gatctgactg atgggctgcc tgtatcgagt ggtgattttg tgccgagctg
      241 ccggtcgggg agctgttggc tggctggtgg caggatatat tgtggtgtaa acaaattgac
      301 gcttagacaa cttaataaca cattgcggac gtttttaatg tactggggtg gatgcagtgg
      361 gccccactct gtgaagacaa actagaattc gagctcggag gtcaacatgg tggagcacga
      421 cactctggtc tactccaaaa atgtcaaaga tacagtctca gaagatcaaa gggctattga
      481 gacttttcaa caaaggataa tttcgggaaa cctcctcgga ttccattgcc cagctatctg
      541 tcacttcatc gaaaggacag tagaaaagga aggtggctcc tacaaatgcc atcattgcga
      601 taaaggaaag gctatcattc aagatctctc tgccgacagt ggtcccaaag atggaccccc
      661 acccacgagg agcatcgtgg aaaaagaaga ggttccaacc acgtctacaa agcaagtgga
      721 ttgatgtgac atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga
      781 cccttcctct atataaggaa gttcatttca tttggagagg acacgctact gtataagagc
      841 tcatttttac aacaattacc aacaacaaca aacaacaaac aacattacaa ttacatttac
      901 aattatcgat aacatggctt cttctatgct ttcttctgct gctgttgttg ctactcgtgc
      961 tagtgctgct caagctagta tggttgctcc ttttactgga cttaagtctg ctgcttcttt
     1021 tcctgttact agaaagcaaa acaaccttga tattacttct attgctagta acggaggaag
     1081 agttcgagca atgggttctc aagaaacaaa tttggctcct catgttctca tattcccttt
     1141 gccaattcaa ggccatgtga actccatgct cagactagca gaacttctct gtctagctga
     1201 attagatatc acttttattg tttcagagtt cagccacagt cgcttaatta agcatacaaa
     1261 tgttgcctcc cgttttgccc gttacccagg atttcagttc cagcccatat ccgatggcct
     1321 tcccgatgat caccccagag caggcgaaag agtcatggat attttgcctt caactaaaaa
     1381 tgtaacggga cccctgttta agcagatgat ggttgagaat aagtgctttt cttctgctac
     1441 tcgaaggccg attacttgca taatcgctga tggggttttg agctttgctg gggatttcgc
     1501 tcaagaaaaa ggaatccctc ttatttactt cagaactgtt tctgcttgtt ctttctgggc
     1561 ttgtttttgt atgcctgaac taattgaatc tggtgacatt ccgattaaag gaaatggtat
     1621 ggatctgata gtgaaaagcg tgccggggat ggaaactttt ctcaggcgac gagatctgcc
     1681 ggggttttgc cgagtcaatg acattaatga accgaaactc cagatcctga aaacagagac
     1741 aagacagaca acgagagccc aagcggctat tttgaacact tttgaggatt tggaaggacc
     1801 aattctatct caaattcgca aacacatgcc aagacttttc acaattggac caagtcattc
     1861 ccacctaaca tctagacttg aaaccaagaa tatcaagact ttaatttcct ccggtagttt
     1921 ctgggaagaa gatcgaagct gtgtggattg gcttgatgca cagccaccaa gatctgtttt
     1981 atatgtaagt tttggaagta ttacagttgt aacaagagat caactcctgg aattctggta
     2041 tggtcttgtt aatagtggac aacggttctt gtgggtaatg aggccagatt caattatggg
     2101 caaagatggg caaagccaga ttcctgcaga tcttgaagag ggcacaaaag ccagaggtta
     2161 tatggtggga tgggcacctc aagaagaggt cttgaatcac cctgccattg gaggatttct
     2221 gactcacagt gggtggaatt caactttaga gagtattgtg gctggtgtcc caatgatatg
     2281 ttggccatac tttgcggacc aaatgataaa cagcagattt gtgagtgaaa tttggaagat
     2341 tggattggat atgaaagata catgtgatag agagacaatt gtgaagatgg ttagggagtt
     2401 gatggagatt aggaaagatg agttcttgca aagggctgat catatggcta aattggcaaa
     2461 agaggctgtt agtgaaggcg gatcctcata ttctaacttg gatggcctcg ttgattacat
     2521 aaaatcactg attatttgag cttctctagc tagagtcgat cgacaagctc gagtttctcc
     2581 ataataatgt gtgagtagtt cccagataag ggaattaggg ttcctatagg gtttcgctca
     2641 tgtgttgagc atataagaaa cccttagtat gtatttgtat ttgtaaaata cttctatcaa
     2701 taaaatttct aattcctaaa accaaaatcc agtactaaaa tccagatcgc tttacttgtc
     2761 ttctgcacga agtggtttaa actatcagtg tttgacagga tatattggcg ggtaaaccta
     2821 agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa aggtttatcc
     2881 gttcgtccat ttgtatgtgc atgccaacca cagggttccc cagatcaggc gctggctgct
     2941 gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac caggtcatca
     3001 ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc ttcgccgacc
     3061 tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg gaaggtttcc
     3121 agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg tcgggccgtc
     3181 ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc agcaaacagc
     3241 acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc acggtccagg
     3301 acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc ggacgtgaag
     3361 cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta ataccggcca
     3421 ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat cggctcgccg
     3481 ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc atcgtcggcc
     3541 cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa aatgaccttg
     3601 ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc agagcgggcc
     3661 gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa caaggaaagc
     3721 tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt ggcctcgctg
     3781 acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat agttcctcgc
     3841 gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcaagacg acgcgaacgc
     3901 tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct gtcgcgctcg
     3961 atcttggccg tagcttgctg gaccatcgag ccgacggact ggaaggtttc gcggggcgca
     4021 cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc cgcgtcgatc
     4081 agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc ttgcgggatt
     4141 gccccgactc acgccggggc aatgtgccct tattcctgat ttgacccgcc tggtgccttg
     4201 gtgtccagat aatccacctt atcggcaatg aagtcggtcc cgtagaccgt ctggccgtcc
     4261 ttctcgtact tggtattccg aatcttgccc tgcacgaata ccagcgaccc cttgcccaaa
     4321 tacttgccgt gggcctcggc ctgagagcca aaacacttga tgcggaagaa gtcggtgcgc
     4381 tcctgcttgt cgccggcatc gttgcgccac atctaggatc tgccaggaac cgtaaaaagg
     4441 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     4501 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     4561 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     4621 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     4681 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     4741 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4801 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4861 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     4921 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     4981 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     5041 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     5101 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     5161 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     5221 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     5281 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     5341 ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca ataaaccagc
     5401 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     5461 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     5521 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     5581 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     5641 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     5701 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     5761 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     5821 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     5881 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     5941 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     6001 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     6061 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat caaggttatt
     6121 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     6181 gcacgaattg gccagcgctg ccatttttgg ggtgaggccg ttcgcggccg aggggcgcag
     6241 cccctggggg gatgggaggc ccgcgttagc gggccgggag ggttcgagaa gggggggcac
     6301 cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa acaaggttta
     6361 taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa aacgggcgga
     6421 aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc aataggtgcg
     6481 cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct catctgtcag
     6541 tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt gtccacatca
     6601 tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct ggccagctcc
     6661 acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg ggtgagtcgg
     6721 ccc
//
