LOCUS       sequence-349519-        6858 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; contains a chloroplast transit peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6858)
  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 6858)
  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: pEPQD1CB0772
FEATURES             Location/Qualifiers
     source          1..6858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    152..176
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        362..707
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     650..670
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2593..2767
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2812..2836
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2992..4140)
                     /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(4476..5064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4556..4575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5235..6026,6027..6095))
                     /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     5858..5877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6096..6200)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(6287..6858,1..60)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tttgcagggc catagacggc cgccagccca gcggcgaggg caaccagccc ggtgagcgtc
       61 gcaaaggaga tcctgatctg actgatgggc tgcctgtatc gagtggtgat tttgtgccga
      121 gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat
      181 tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactgg ggtggatgca
      241 gtgggcccca ctctgtgaag acaatgtgga attcctcgag ggaggtcaac atggtggagc
      301 acgacactct ggtctactcc aaaaatgtca aagatacagt ctcagaagat caaagggcta
      361 ttgagacttt tcaacaaagg ataatttcgg gaaacctcct cggattccat tgcccagcta
      421 tctgtcactt catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt
      481 gcgataaagg aaaggctatc attcaagatc tctctgccga cagtggtccc aaagatggac
      541 ccccacccac gaggagcatc gtggaaaaag aagaggttcc aaccacgtct acaaagcaag
      601 tggattgatg tgacatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc
      661 aagacccttc ctctatataa ggaagttcat ttcatttgga gaggacacgc tactgtataa
      721 gagctcattt ttacaacaat taccaacaac aacaaacaac aaacaacatt acaattacat
      781 ttacaattat cgataacatg gcttcttcta tgctttcttc tgctgctgtt gttgctactc
      841 gtgctagtgc tgctcaagct agtatggttg ctccttttac tggacttaag tctgctgctt
      901 cttttcctgt tactagaaag caaaacaacc ttgatattac ttctattgct agtaacggag
      961 gaagagttcg agcaatggag atggatatgg atatcattag aaaggcaatt gctgccacta
     1021 tctttgcatt ggttatggct tgggcgtgga gagtgttgga ttgggcatgg tttactccta
     1081 agaggatcga gaaacgtcta aggcagcaag gttttagagg aaatccttat agattcttgg
     1141 ttggagatgt taaggagagt ggaaaaatgc atcaagaagc cttgtctaac cccatggagt
     1201 tcgacaacga tattgttcct cgcctcatgc cacatattaa ccacaccata aaaacttacg
     1261 gtaggaattc atttacatgg atgggaagga ttccaaggat tcatgttatg gaacctgaac
     1321 ttattaagga agtattgacc cactccagca aataccaaaa gaactttgat gttcacaatc
     1381 cccttgttaa gttccttctc actggagttg gaagctttga aggggcaaaa tggtccaaac
     1441 acagaagaat catttcccct gccttcactc ttgagaaact aaagtcaatg ttgccggctt
     1501 ttgccatatg ctatcatgac atgttgacga aatgggagaa attagcagaa aaagaaggat
     1561 cccatgaagt tgatatcttc cccacatttg atgttttgac aagtgatgta atttcaaagg
     1621 ttgcatttgg tagcacatac gatgaaggag gcaaaatctt cagactattg aaagaactca
     1681 tggatctcac aattgattgc atgagagatg tctacatccc agggtggagc tacttgccaa
     1741 ccaagaggaa caagaggatg aaagaaatta acaaagaaat cacagatatg ctaaggttca
     1801 tcatcaacaa gagaatgaag gctttgaagg ctggagaacc aggtgaggat gacttgttgg
     1861 gagtcctgtt ggaatcaaac attcaagaaa ttcagaaaca aggaaacagg aaggatggtg
     1921 gaatgacaat caacgatgtg attgaggaat gcaaattgtt ctactttgct ggtcaagaaa
     1981 ccaccggagt tttgctcaca tggaccacca ttttattgag caagcatccc gagtggcaag
     2041 agcgagctag ggaagaagtt ctccaagcct ttggcaagaa caagcctgaa tttgaacgct
     2101 tgaatcatct taaatatgtt tctatgatct tatacgaggt tctaaggttg tacccgccag
     2161 ttattgatct gaccaagatt attcacgagg acacaaagtt agggccgtac acaatccctg
     2221 caggaacaca agtaatgttg ccgacagtaa tgcttcatag agagaagtcc atttggggag
     2281 aagatgcaat ggaattcaac ccaatgagat ttgctgatgg agttgccaat gccaccaaga
     2341 acaatgtcac atatttgcca ttcagttggg gacctagagt ttgcctcggc caaaactttg
     2401 cacttcttca agcaaaatta gggttggcaa tgattttaca acgcttcaaa tttgatgtag
     2461 ctccatccta tgttcatgct ccctttacca ttcttacagt tcaacctcag tttggttctc
     2521 atgttatcta caagaagctt gagagacaga atttttaagc ttctctagct agagtcgatc
     2581 gacaagctcg agtttctcca taataatgtg tgagtagttc ccagataagg gaattagggt
     2641 tcctataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt
     2701 tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtactaaaat
     2761 ccagatcgct gagcttgtct tctgcacgaa gtggtttaaa ctatcagtgt ttgacaggat
     2821 atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag
     2881 ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc
     2941 agatcaggcg ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt
     3001 gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc
     3061 ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca
     3121 catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc
     3181 gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta
     3241 gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt
     3301 tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc
     3361 aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc
     3421 cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt
     3481 gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac
     3541 cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt
     3601 gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt
     3661 gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc
     3721 aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc
     3781 ggcctgcttg gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt
     3841 ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg
     3901 ttcaagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag
     3961 ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg
     4021 gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc
     4081 ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat
     4141 tcaccctcct tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt
     4201 tgacccgcct ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc
     4261 gtagaccgtc tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac
     4321 cagcgacccc ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa aacacttgat
     4381 gcggaagaag tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca tctaggatct
     4441 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4501 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4561 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4621 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4681 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4741 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4801 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4861 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4921 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4981 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5041 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5101 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5161 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5221 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5281 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5341 ttaccatctg gccccagtgc tgcaatgata ccgcgagaac cacgctcacc ggctccagat
     5401 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5461 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5521 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5581 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5641 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5701 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5761 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5821 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5881 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5941 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6001 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6061 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6121 agcatttatc aaggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6181 aaacaaatag gggttccgcg cacgaattgg ccagcgctgc catttttggg gtgaggccgt
     6241 tcgcggccga ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg ggccgggagg
     6301 gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag ggcgcagccc
     6361 tggttaaaaa caaggtttat aaatattggt ttaaaagcag gttaaaagac aggttagcgg
     6421 tggccgaaaa acgggcggaa acccttgcaa atgctggatt ttctgcctgt ggacagcccc
     6481 tcaaatgtca ataggtgcgc ccctcatctg tcagcactct gcccctcaag tgtcaaggat
     6541 cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag ggcacttatc
     6601 cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt ttcgccgatt
     6661 tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag cctgcccctc atctgtcaac
     6721 gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct gtcagtgagg
     6781 gccaagtttt ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc acacggcttc
     6841 gacggcgttt ctggcgcg
//
