LOCUS       sequence-349526-        6333 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 2, transcriptional unit for transient 
            expression of strictosidine synthase (CrSTR) 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 6333)
  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 6333)
  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: pEPQD1CB0776
FEATURES             Location/Qualifiers
     source          1..6333
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    183..207
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        393..738
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     681..701
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     polyA_signal    2099..2273
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2318..2342
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2498..3646)
                     /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(3982..4570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4062..4081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4741..5532,5533..5601))
                     /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     5364..5383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5602..5706)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5793..6333,1..91)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 cgcacacggc ttcgacggcg tttctggcgc gtttgcaggg ccatagacgg ccgccagccc
       61 agcggcgagg gcaaccagcc cggtgagcgt cgcaaaggag atcctgatct gactgatggg
      121 ctgcctgtat cgagtggtga ttttgtgccg agctgccggt cggggagctg ttggctggct
      181 ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta gacaacttaa taacacattg
      241 cggacgtttt taatgtactg gggtggatgc agtgggcccc actctgtgaa gacaagcaag
      301 aattcaagct tggaggtcaa catggtggag cacgacactc tggtctactc caaaaatgtc
      361 aaagatacag tctcagaaga tcaaagggct attgagactt ttcaacaaag gataatttcg
      421 ggaaacctcc tcggattcca ttgcccagct atctgtcact tcatcgaaag gacagtagaa
      481 aaggaaggtg gctcctacaa atgccatcat tgcgataaag gaaaggctat cattcaagat
      541 ctctctgccg acagtggtcc caaagatgga cccccaccca cgaggagcat cgtggaaaaa
      601 gaagaggttc caaccacgtc tacaaagcaa gtggattgat gtgacatctc cactgacgta
      661 agggatgacg cacaatccca ctatccttcg caagaccctt cctctatata aggaagttca
      721 tttcatttgg agaggacacg ctactgtata agagctcatt tttacaacaa ttaccaacaa
      781 caacaaacaa caaacaacat tacaattaca tttacaatta tcgataacat ggcttcttct
      841 atgctttctt ctgctgctgt tgttgctact cgtgctagtg ctgctcaagc tagtatggtt
      901 gctcctttta ctggacttaa gtctgctgct tcttttcctg ttactagaaa gcaaaacaac
      961 cttgatatta cttctattgc tagtaacgga ggaagagttc gagcaatggc aaacttttct
     1021 gaatctaaat ccatgatggc agttttcttc atgtttttcc tgcttctttt gtcatcttct
     1081 agttcttcat cttcaagttc accaattttg aaaaagattt ttattgaaag cccttcctat
     1141 gctccgaatg ccttcacctt cgattcaact gataaagggt tctacacttc cgtccaagat
     1201 ggccgagtta tcaaatatga agggccaaat tcaggcttca ctgacttcgc ctacgcatct
     1261 cccttctgga acaaagcttt ttgtgagaac agcaccgatc cagagaaaag accattgtgt
     1321 gggaggacat atgatatttc ctatgactat aagaacagcc aaatgtacat tgttgatggc
     1381 cattaccatc tttgtgtggt tggaaaagaa ggtgggtatg ccacacaact agccacaagt
     1441 gtgcaaggag tgccattcaa atggctctat gcagtaactg ttgatcagag aacagggatt
     1501 gtttatttca ctgatgttag ctccatacat gatgacagtc ccgaaggtgt ggaagaaatc
     1561 atgaatacaa gtgatagaac agggagatta atgaagtatg atccttcaac aaaagaaacc
     1621 accttattat tgaaagagct acatgttccc ggcggtgcag aaatcagcgc agatggttcc
     1681 tttgttgtag tagcagaatt tttaagcaat cggatagtga agtattggct agaagggcca
     1741 aagaaaggca gtgcagagtt cttagttaca atcccaaatc caggaaatat aaagaggaat
     1801 tctgatggcc atttttgggt gtcatctagt gaagaattag atggaggtca acatggaaga
     1861 gttgtttcaa gaggaattaa gtttgatgga tttgggaata ttcttcaagt tataccactt
     1921 ccaccaccat atgaaggtga acattttgaa cagattcaag agcacgatgg tttgttatac
     1981 attggaagtc tcttccatag ctctgtgggt atattagtgt atgatgatca tgataacaag
     2041 ggaaattctt atgtttctag ctgagcttct ctagctagag tcgatcgaca agctcgagtt
     2101 tctccataat aatgtgtgag tagttcccag ataagggaat tagggttcct atagggtttc
     2161 gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct
     2221 atcaataaaa tttctaattc ctaaaaccaa aatccagtac taaaatccag atcgctacta
     2281 ttgtcttctg cacgaagtgg tttaaactat cagtgtttga caggatatat tggcgggtaa
     2341 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
     2401 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttccccagat caggcgctgg
     2461 ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt
     2521 catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc
     2581 cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg
     2641 tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg
     2701 ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa
     2761 acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt
     2821 ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg
     2881 tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc
     2941 ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct
     3001 cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt
     3061 cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga
     3121 ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc
     3181 gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg
     3241 aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct
     3301 cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc
     3361 ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttca agacgacgcg
     3421 aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc
     3481 gctcgatctt ggccgtagct tgctggacca tcgagccgac ggactggaag gtttcgcggg
     3541 gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt
     3601 cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg
     3661 ggattgcccc gactcacgcc ggggcaatgt gcccttattc ctgatttgac ccgcctggtg
     3721 ccttggtgtc cagataatcc accttatcgg caatgaagtc ggtcccgtag accgtctggc
     3781 cgtccttctc gtacttggta ttccgaatct tgccctgcac gaataccagc gaccccttgc
     3841 ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg
     3901 tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa
     3961 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4021 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4081 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4141 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4201 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4261 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4321 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4381 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4441 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4501 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4561 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4621 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4681 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4741 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4801 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4861 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     4921 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4981 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5041 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5101 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5161 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5221 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5281 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5341 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5401 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5461 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5521 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5581 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaagg
     5641 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5701 tccgcgcacg aattggccag cgctgccatt tttggggtga ggccgttcgc ggccgagggg
     5761 cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg
     5821 ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag
     5881 gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg
     5941 gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag
     6001 gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct
     6061 gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca
     6121 catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca
     6181 gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga
     6241 gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc
     6301 gaggtatcca caacgccggc ggccgcggtg tct
//
