LOCUS       sequence-349498-        6441 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; contains a chloroplast transit peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6441)
  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 6441)
  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: pEPQD1CB0118
FEATURES             Location/Qualifiers
     source          1..6441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(47..838,839..907))
                     /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     670..689
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(908..1012)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1099..1730
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    1822..1846
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        2032..2377
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     2320..2340
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    3846..4020
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    4065..4089
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4245..5393)
                     /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(5729..6317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5809..5828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
       61 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      121 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      181 taccgcgaga accacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
      241 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
      301 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
      361 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
      421 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
      481 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
      541 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
      601 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
      661 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
      721 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
      781 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
      841 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
      901 tactcatact cttccttttt caatattatt gaagcattta tcaaggttat tgtctcatga
      961 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacgaatt
     1021 ggccagcgct gccatttttg gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg
     1081 ggatgggagg cccgcgttag cgggccggga gggttcgaga agggggggca ccccccttcg
     1141 gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa aacaaggttt ataaatattg
     1201 gtttaaaagc aggttaaaag acaggttagc ggtggccgaa aaacgggcgg aaacccttgc
     1261 aaatgctgga ttttctgcct gtggacagcc cctcaaatgt caataggtgc gcccctcatc
     1321 tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc
     1381 ccctcaagtg tcaataccgc agggcactta tccccaggct tgtccacatc atctgtggga
     1441 aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc tggccagctc cacgtcgccg
     1501 gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag
     1561 tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt ttccgcgagg tatccacaac
     1621 gccggcggcc gcggtgtctc gcacacggct tcgacggcgt ttctggcgcg tttgcagggc
     1681 catagacggc cgccagccca gcggcgaggg caaccagccc ggtgagcgtc gcaaaggaga
     1741 tcctgatctg actgatgggc tgcctgtatc gagtggtgat tttgtgccga gctgccggtc
     1801 ggggagctgt tggctggctg gtggcaggat atattgtggt gtaaacaaat tgacgcttag
     1861 acaacttaat aacacattgc ggacgttttt aatgtactgg ggtggatgca gtgggcccca
     1921 ctctgtgaag acaatgtgga attcctcgag ggaggtcaac atggtggagc acgacactct
     1981 ggtctactcc aaaaatgtca aagatacagt ctcagaagat caaagggcta ttgagacttt
     2041 tcaacaaagg ataatttcgg gaaacctcct cggattccat tgcccagcta tctgtcactt
     2101 catcgaaagg acagtagaaa aggaaggtgg ctcctacaaa tgccatcatt gcgataaagg
     2161 aaaggctatc attcaagatc tctctgccga cagtggtccc aaagatggac ccccacccac
     2221 gaggagcatc gtggaaaaag aagaggttcc aaccacgtct acaaagcaag tggattgatg
     2281 tgacatctcc actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc
     2341 ctctatataa ggaagttcat ttcatttgga gaggacacgc tactgtataa gagctcattt
     2401 ttacaacaat taccaacaac aacaaacaac aaacaacatt acaattacat ttacaattat
     2461 cgataacatg gcttcttcta tgctttcttc tgctgctgtt gttgctactc gtgctagtgc
     2521 tgctcaagct agtatggttg ctccttttac tggacttaag tctgctgctt cttttcctgt
     2581 tactagaaag caaaacaacc ttgatattac ttctattgct agtaacggag gaagagttcg
     2641 agcaatgagt tggtggtgga agaggtccat tggtgctgga aagaacttac caaaccagaa
     2701 caaagaaaat ggagtttgta aaagctacaa gagtgtggca ctagtagtag gagtcaccgg
     2761 cattgttggc agtagtttag ccgaagttct aaagctacca gacacccctg gaggtccctg
     2821 gaaggtatac ggcgtggcaa ggcgcccatg ccctgtatgg ctagccaaaa aaccagttga
     2881 gtacatccaa tgtgatgtct cagataatca agaaacaatt tctaaattat ctccacttaa
     2941 ggacataact catatattct atgtttcttg gattggatca gaagattgcc aaacaaatgc
     3001 tacaatgttc aaaaacatcc ttaattcagt gattcctaat gcttcaaatc tccaacatgt
     3061 gtgtctccaa acaggaatca aacattattt tggtattttt gaagaaggtt ctaaagttgt
     3121 accacatgat tcgcctttta ccgaggattt acctaggcta aatgtcccaa atttctacca
     3181 tgatcttgaa gatatactgt atgaagaaac tggcaagaac aatttaacat ggtctgtcca
     3241 tagaccagct ctggttttcg ggttttcccc atgtagtatg atgaacattg tcagtacatt
     3301 atgcgtttac gccacaattt gtaaacatga aaataaggcc ttggtttatc caggtagtaa
     3361 aaattcatgg aattgttatg ctgatgctgt tgatgcagat ttagtagccg agcacgaaat
     3421 ttgggcagca gttgatccta aggctaaaaa tcaagtattg aattgtaaca atggggatgt
     3481 ttttaaatgg aaacatattt ggaagaaatt agcagaggaa tttgggattg aaatggtggg
     3541 atatgttgaa ggtaaagaac aagttagttt ggctgagtta atgaaggata aagatcaagt
     3601 ttgggatgaa attgttaaga agaacaattt agtacctact aaattgaaag aaattgctgc
     3661 cttttggttt gctgatattg ctttttgttc tgagaatttg attagtagta tgaacaaaag
     3721 taaggaatta gggttcttgg gttttaggaa ttcgatgaaa tcgtttgttt cttgtattga
     3781 taagatgagg gattataggt ttattcctta ggcttctcta gctagagtcg atcgacaagc
     3841 tcgagtttct ccataataat gtgtgagtag ttcccagata agggaattag ggttcctata
     3901 gggtttcgct catgtgttga gcatataaga aacccttagt atgtatttgt atttgtaaaa
     3961 tacttctatc aataaaattt ctaattccta aaaccaaaat ccagtactaa aatccagatc
     4021 gctgagcttg tcttctgcac gaagtggttt aaactatcag tgtttgacag gatatattgg
     4081 cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa aagggcgtga
     4141 aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc cccagatcag
     4201 gcgctggctg ctgaaccccc agccggaact gaccccacaa ggccctagcg tttgcaatgc
     4261 accaggtcat cattgaccca ggcgtgttcc accaggccgc tgcctcgcaa ctcttcgcag
     4321 gcttcgccga cctgctcgcg ccacttcttc acgcgggtgg aatccgatcc gcacatgagg
     4381 cggaaggttt ccagcttgag cgggtacggc tcccggtgcg agctgaaata gtcgaacatc
     4441 cgtcgggccg tcggcgacag cttgcggtac ttctcccata tgaatttcgt gtagtggtcg
     4501 ccagcaaaca gcacgacgat ttcctcgtcg atcaggacct ggcaacggga cgttttcttg
     4561 ccacggtcca ggacgcggaa gcggtgcagc agcgacaccg attccaggtg cccaacgcgg
     4621 tcggacgtga agcccatcgc cgtcgcctgt aggcgcgaca ggcattcctc ggccttcgtg
     4681 taataccggc cattgatcga ccagcccagg tcctggcaaa gctcgtagaa cgtgaaggtg
     4741 atcggctcgc cgataggggt gcgcttcgcg tactccaaca cctgctgcca caccagttcg
     4801 tcatcgtcgg cccgcagctc gacgccggtg taggtgatct tcacgtcctt gttgacgtgg
     4861 aaaatgacct tgttttgcag cgcctcgcgc gggattttct tgttgcgcgt ggtgaacagg
     4921 gcagagcggg ccgtgtcgtt tggcatcgct cgcatcgtgt ccggccacgg cgcaatatcg
     4981 aacaaggaaa gctgcatttc cttgatctgc tgcttcgtgt gtttcagcaa cgcggcctgc
     5041 ttggcctcgc tgacctgttt tgccaggtcc tcgccggcgg tttttcgctt cttggtcgtc
     5101 atagttcctc gcgtgtcgat ggtcatcgac ttcgccaaac ctgccgcctc ctgttcaaga
     5161 cgacgcgaac gctccacggc ggccgatggc gcgggcaggg cagggggagc cagttgcacg
     5221 ctgtcgcgct cgatcttggc cgtagcttgc tggaccatcg agccgacgga ctggaaggtt
     5281 tcgcggggcg cacgcatgac ggtgcggctt gcgatggttt cggcatcctc ggcggaaaac
     5341 cccgcgtcga tcagttcttg cctgtatgcc ttccggtcaa acgtccgatt cattcaccct
     5401 ccttgcggga ttgccccgac tcacgccggg gcaatgtgcc cttattcctg atttgacccg
     5461 cctggtgcct tggtgtccag ataatccacc ttatcggcaa tgaagtcggt cccgtagacc
     5521 gtctggccgt ccttctcgta cttggtattc cgaatcttgc cctgcacgaa taccagcgac
     5581 cccttgccca aatacttgcc gtgggcctcg gcctgagagc caaaacactt gatgcggaag
     5641 aagtcggtgc gctcctgctt gtcgccggca tcgttgcgcc acatctagga tctgccagga
     5701 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     5761 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     5821 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5881 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5941 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     6001 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     6061 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     6121 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     6181 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     6241 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     6301 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     6361 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     6421 tccttttaaa ttaaaaatga a
//
