LOCUS       sequence-349518-        6681 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 6, transcriptional unit for transient 
            expression of secologanin synthase (CrSLS1) from Catharanthus roseus
            driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6681)
  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 6681)
  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: pEPQD1CB0771
FEATURES             Location/Qualifiers
     source          1..6681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    168..192
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        378..723
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     666..686
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2432..2606
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2651..2675
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2831..3979)
                     /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(4315..4903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4395..4414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5074..5865,5866..5934))
                     /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     5697..5716
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5935..6039)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6126..6681,1..76)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cggcgtttct ggcgcgtttg cagggccata gacggccgcc agcccagcgg cgagggcaac
       61 cagcccggtg agcgtcgcaa aggagatcct gatctgactg atgggctgcc tgtatcgagt
      121 ggtgattttg tgccgagctg ccggtcgggg agctgttggc tggctggtgg caggatatat
      181 tgtggtgtaa acaaattgac gcttagacaa cttaataaca cattgcggac gtttttaatg
      241 tactggggtg gatgcagtgg gccccactct gtgaagacaa tgtggaattc ctcgagggag
      301 gtcaacatgg tggagcacga cactctggtc tactccaaaa atgtcaaaga tacagtctca
      361 gaagatcaaa gggctattga gacttttcaa caaaggataa tttcgggaaa cctcctcgga
      421 ttccattgcc cagctatctg tcacttcatc gaaaggacag tagaaaagga aggtggctcc
      481 tacaaatgcc atcattgcga taaaggaaag gctatcattc aagatctctc tgccgacagt
      541 ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga ggttccaacc
      601 acgtctacaa agcaagtgga ttgatgtgac atctccactg acgtaaggga tgacgcacaa
      661 tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca tttggagagg
      721 acacgctcga gtataagagc tcatttttac aacaattacc aacaacaaca aacaacaaac
      781 aacattacaa ttacatttac aattatcgat acaatggaga tggatatgga tatcattaga
      841 aaggcaattg ctgccactat ctttgcattg gttatggctt gggcgtggag agtgttggat
      901 tgggcatggt ttactcctaa gaggatcgag aaacgtctaa ggcagcaagg ttttagagga
      961 aatccttata gattcttggt tggagatgtt aaggagagtg gaaaaatgca tcaagaagcc
     1021 ttgtctaacc ccatggagtt cgacaacgat attgttcctc gcctcatgcc acatattaac
     1081 cacaccataa aaacttacgg taggaattca tttacatgga tgggaaggat tccaaggatt
     1141 catgttatgg aacctgaact tattaaggaa gtattgaccc actccagcaa ataccaaaag
     1201 aactttgatg ttcacaatcc ccttgttaag ttccttctca ctggagttgg aagctttgaa
     1261 ggggcaaaat ggtccaaaca cagaagaatc atttcccctg ccttcactct tgagaaacta
     1321 aagtcaatgt tgccggcttt tgccatatgc tatcatgaca tgttgacgaa atgggagaaa
     1381 ttagcagaaa aagaaggatc ccatgaagtt gatatcttcc ccacatttga tgttttgaca
     1441 agtgatgtaa tttcaaaggt tgcatttggt agcacatacg atgaaggagg caaaatcttc
     1501 agactattga aagaactcat ggatctcaca attgattgca tgagagatgt ctacatccca
     1561 gggtggagct acttgccaac caagaggaac aagaggatga aagaaattaa caaagaaatc
     1621 acagatatgc taaggttcat catcaacaag agaatgaagg ctttgaaggc tggagaacca
     1681 ggtgaggatg acttgttggg agtcctgttg gaatcaaaca ttcaagaaat tcagaaacaa
     1741 ggaaacagga aggatggtgg aatgacaatc aacgatgtga ttgaggaatg caaattgttc
     1801 tactttgctg gtcaagaaac caccggagtt ttgctcacat ggaccaccat tttattgagc
     1861 aagcatcccg agtggcaaga gcgagctagg gaagaagttc tccaagcctt tggcaagaac
     1921 aagcctgaat ttgaacgctt gaatcatctt aaatatgttt ctatgatctt atacgaggtt
     1981 ctaaggttgt acccgccagt tattgatctg accaagatta ttcacgagga cacaaagtta
     2041 gggccgtaca caatccctgc aggaacacaa gtaatgttgc cgacagtaat gcttcataga
     2101 gagaagtcca tttggggaga agatgcaatg gaattcaacc caatgagatt tgctgatgga
     2161 gttgccaatg ccaccaagaa caatgtcaca tatttgccat tcagttgggg acctagagtt
     2221 tgcctcggcc aaaactttgc acttcttcaa gcaaaattag ggttggcaat gattttacaa
     2281 cgcttcaaat ttgatgtagc tccatcctat gttcatgctc cctttaccat tcttacagtt
     2341 caacctcagt ttggttctca tgttatctac aagaagcttg agagacagaa tttttaagct
     2401 tctctagcta gagtcgatcg acaagctcga gtttctccat aataatgtgt gagtagttcc
     2461 cagataaggg aattagggtt cctatagggt ttcgctcatg tgttgagcat ataagaaacc
     2521 cttagtatgt atttgtattt gtaaaatact tctatcaata aaatttctaa ttcctaaaac
     2581 caaaatccag tactaaaatc cagatcgctg agcttgtctt ctgcacgaag tggtttaaac
     2641 tatcagtgtt tgacaggata tattggcggg taaacctaag agaaaagagc gtttattaga
     2701 ataatcggat atttaaaagg gcgtgaaaag gtttatccgt tcgtccattt gtatgtgcat
     2761 gccaaccaca gggttcccca gatcaggcgc tggctgctga acccccagcc ggaactgacc
     2821 ccacaaggcc ctagcgtttg caatgcacca ggtcatcatt gacccaggcg tgttccacca
     2881 ggccgctgcc tcgcaactct tcgcaggctt cgccgacctg ctcgcgccac ttcttcacgc
     2941 gggtggaatc cgatccgcac atgaggcgga aggtttccag cttgagcggg tacggctccc
     3001 ggtgcgagct gaaatagtcg aacatccgtc gggccgtcgg cgacagcttg cggtacttct
     3061 cccatatgaa tttcgtgtag tggtcgccag caaacagcac gacgatttcc tcgtcgatca
     3121 ggacctggca acgggacgtt ttcttgccac ggtccaggac gcggaagcgg tgcagcagcg
     3181 acaccgattc caggtgccca acgcggtcgg acgtgaagcc catcgccgtc gcctgtaggc
     3241 gcgacaggca ttcctcggcc ttcgtgtaat accggccatt gatcgaccag cccaggtcct
     3301 ggcaaagctc gtagaacgtg aaggtgatcg gctcgccgat aggggtgcgc ttcgcgtact
     3361 ccaacacctg ctgccacacc agttcgtcat cgtcggcccg cagctcgacg ccggtgtagg
     3421 tgatcttcac gtccttgttg acgtggaaaa tgaccttgtt ttgcagcgcc tcgcgcggga
     3481 ttttcttgtt gcgcgtggtg aacagggcag agcgggccgt gtcgtttggc atcgctcgca
     3541 tcgtgtccgg ccacggcgca atatcgaaca aggaaagctg catttccttg atctgctgct
     3601 tcgtgtgttt cagcaacgcg gcctgcttgg cctcgctgac ctgttttgcc aggtcctcgc
     3661 cggcggtttt tcgcttcttg gtcgtcatag ttcctcgcgt gtcgatggtc atcgacttcg
     3721 ccaaacctgc cgcctcctgt tcaagacgac gcgaacgctc cacggcggcc gatggcgcgg
     3781 gcagggcagg gggagccagt tgcacgctgt cgcgctcgat cttggccgta gcttgctgga
     3841 ccatcgagcc gacggactgg aaggtttcgc ggggcgcacg catgacggtg cggcttgcga
     3901 tggtttcggc atcctcggcg gaaaaccccg cgtcgatcag ttcttgcctg tatgccttcc
     3961 ggtcaaacgt ccgattcatt caccctcctt gcgggattgc cccgactcac gccggggcaa
     4021 tgtgccctta ttcctgattt gacccgcctg gtgccttggt gtccagataa tccaccttat
     4081 cggcaatgaa gtcggtcccg tagaccgtct ggccgtcctt ctcgtacttg gtattccgaa
     4141 tcttgccctg cacgaatacc agcgacccct tgcccaaata cttgccgtgg gcctcggcct
     4201 gagagccaaa acacttgatg cggaagaagt cggtgcgctc ctgcttgtcg ccggcatcgt
     4261 tgcgccacat ctaggatctg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4321 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4381 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4441 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4501 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4561 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4621 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4681 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     4741 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4801 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4861 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4921 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     4981 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     5041 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     5101 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     5161 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc
     5221 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     5281 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     5341 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     5401 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     5461 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     5521 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     5581 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     5641 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     5701 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     5761 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     5821 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     5881 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     5941 cctttttcaa tattattgaa gcatttatca aggttattgt ctcatgagcg gatacatatt
     6001 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acgaattggc cagcgctgcc
     6061 atttttgggg tgaggccgtt cgcggccgag gggcgcagcc cctgggggga tgggaggccc
     6121 gcgttagcgg gccgggaggg ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca
     6181 cgcgcacagg gcgcagccct ggttaaaaac aaggtttata aatattggtt taaaagcagg
     6241 ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt
     6301 tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc cctcatctgt cagcactctg
     6361 cccctcaagt gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca
     6421 ataccgcagg gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa
     6481 tcaggcgttt tcgccgattt gcgaggctgg ccagctccac gtcgccggcc gaaatcgagc
     6541 ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt caacgtccgc
     6601 ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc ggcggccgcg
     6661 gtgtctcgca cacggcttcg a
//
