LOCUS       sequence-349497-        6264 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 6, transcriptional unit for transient 
            expression of iridoid synthase (CrISY) from Catharanthus roseus 
            driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6264)
  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 6264)
  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: pEPQD1CB0117
FEATURES             Location/Qualifiers
     source          1..6264
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    136..160
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        346..691
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     634..654
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    1983..2157
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2202..2226
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2382..3530)
                     /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(3866..4454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3946..3965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4625..5416,5417..5485))
                     /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     5248..5267
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5486..5590)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5677..6264,1..44)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cggccgccag cccagcggcg agggcaacca gcccggtgag cgtcgcaaag gagatcctga
       61 tctgactgat gggctgcctg tatcgagtgg tgattttgtg ccgagctgcc ggtcggggag
      121 ctgttggctg gctggtggca ggatatattg tggtgtaaac aaattgacgc ttagacaact
      181 taataacaca ttgcggacgt ttttaatgta ctggggtgga tgcagtgggc cccactctgt
      241 gaagacaatg tggaattcct cgagggaggt caacatggtg gagcacgaca ctctggtcta
      301 ctccaaaaat gtcaaagata cagtctcaga agatcaaagg gctattgaga cttttcaaca
      361 aaggataatt tcgggaaacc tcctcggatt ccattgccca gctatctgtc acttcatcga
      421 aaggacagta gaaaaggaag gtggctccta caaatgccat cattgcgata aaggaaaggc
      481 tatcattcaa gatctctctg ccgacagtgg tcccaaagat ggacccccac ccacgaggag
      541 catcgtggaa aaagaagagg ttccaaccac gtctacaaag caagtggatt gatgtgacat
      601 ctccactgac gtaagggatg acgcacaatc ccactatcct tcgcaagacc cttcctctat
      661 ataaggaagt tcatttcatt tggagaggac acgctcgagt ataagagctc atttttacaa
      721 caattaccaa caacaacaaa caacaaacaa cattacaatt acatttacaa ttatcgatac
      781 aatgagttgg tggtggaaga ggtccattgg tgctggaaag aacttaccaa accagaacaa
      841 agaaaatgga gtttgtaaaa gctacaagag tgtggcacta gtagtaggag tcaccggcat
      901 tgttggcagt agtttagccg aagttctaaa gctaccagac acccctggag gtccctggaa
      961 ggtatacggc gtggcaaggc gcccatgccc tgtatggcta gccaaaaaac cagttgagta
     1021 catccaatgt gatgtctcag ataatcaaga aacaatttct aaattatctc cacttaagga
     1081 cataactcat atattctatg tttcttggat tggatcagaa gattgccaaa caaatgctac
     1141 aatgttcaaa aacatcctta attcagtgat tcctaatgct tcaaatctcc aacatgtgtg
     1201 tctccaaaca ggaatcaaac attattttgg tatttttgaa gaaggttcta aagttgtacc
     1261 acatgattcg ccttttaccg aggatttacc taggctaaat gtcccaaatt tctaccatga
     1321 tcttgaagat atactgtatg aagaaactgg caagaacaat ttaacatggt ctgtccatag
     1381 accagctctg gttttcgggt tttccccatg tagtatgatg aacattgtca gtacattatg
     1441 cgtttacgcc acaatttgta aacatgaaaa taaggccttg gtttatccag gtagtaaaaa
     1501 ttcatggaat tgttatgctg atgctgttga tgcagattta gtagccgagc acgaaatttg
     1561 ggcagcagtt gatcctaagg ctaaaaatca agtattgaat tgtaacaatg gggatgtttt
     1621 taaatggaaa catatttgga agaaattagc agaggaattt gggattgaaa tggtgggata
     1681 tgttgaaggt aaagaacaag ttagtttggc tgagttaatg aaggataaag atcaagtttg
     1741 ggatgaaatt gttaagaaga acaatttagt acctactaaa ttgaaagaaa ttgctgcctt
     1801 ttggtttgct gatattgctt tttgttctga gaatttgatt agtagtatga acaaaagtaa
     1861 ggaattaggg ttcttgggtt ttaggaattc gatgaaatcg tttgtttctt gtattgataa
     1921 gatgagggat tataggttta ttccttaggc ttctctagct agagtcgatc gacaagctcg
     1981 agtttctcca taataatgtg tgagtagttc ccagataagg gaattagggt tcctataggg
     2041 tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt tgtaaaatac
     2101 ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtactaaaat ccagatcgct
     2161 gagcttgtct tctgcacgaa gtggtttaaa ctatcagtgt ttgacaggat atattggcgg
     2221 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa
     2281 ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc agatcaggcg
     2341 ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt gcaatgcacc
     2401 aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc ttcgcaggct
     2461 tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca catgaggcgg
     2521 aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc gaacatccgt
     2581 cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta gtggtcgcca
     2641 gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt tttcttgcca
     2701 cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc aacgcggtcg
     2761 gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc cttcgtgtaa
     2821 taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt gaaggtgatc
     2881 ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac cagttcgtca
     2941 tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt gacgtggaaa
     3001 atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt gaacagggca
     3061 gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc aatatcgaac
     3121 aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc ggcctgcttg
     3181 gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt ggtcgtcata
     3241 gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg ttcaagacga
     3301 cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag ttgcacgctg
     3361 tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg gaaggtttcg
     3421 cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc ggaaaacccc
     3481 gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat tcaccctcct
     3541 tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt tgacccgcct
     3601 ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc gtagaccgtc
     3661 tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac cagcgacccc
     3721 ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa aacacttgat gcggaagaag
     3781 tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca tctaggatct gccaggaacc
     3841 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3901 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3961 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4021 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4081 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4141 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4201 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4261 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4321 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4381 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4441 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4501 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4561 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4621 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4681 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4741 gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat ttatcagcaa
     4801 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4861 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4921 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4981 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5041 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5101 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5161 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5221 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5281 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5341 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5401 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5461 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5521 aaggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5581 gggttccgcg cacgaattgg ccagcgctgc catttttggg gtgaggccgt tcgcggccga
     5641 ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg ggccgggagg gttcgagaag
     5701 ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc tggttaaaaa
     5761 caaggtttat aaatattggt ttaaaagcag gttaaaagac aggttagcgg tggccgaaaa
     5821 acgggcggaa acccttgcaa atgctggatt ttctgcctgt ggacagcccc tcaaatgtca
     5881 ataggtgcgc ccctcatctg tcagcactct gcccctcaag tgtcaaggat cgcgcccctc
     5941 atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc cccaggcttg
     6001 tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt tgcgaggctg
     6061 gccagctcca cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac gccgcgccgg
     6121 gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg gccaagtttt
     6181 ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc acacggcttc gacggcgttt
     6241 ctggcgcgtt tgcagggcca taga
//
