LOCUS       sequence-349542-        6659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 5, transcriptional unit for transient 
            expression of secologanin synthase (CrSLS1) from Catharanthus roseus
            promoter regions contains 4x attB sites for recruitment fo 
            gal4AD-phiC31.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6659)
  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 6659)
  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: pEPQD1CB0863
FEATURES             Location/Qualifiers
     source          1..6659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    89..144
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      1742..1994
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2042..2066
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2222..3370)
                     /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(3706..4294)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3786..3805)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4465..5256,5257..5325))
                     /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     5088..5107
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5326..5430)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5517..6148
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    6240..6264
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    6381..6414
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    6440..6473
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    6503..6536
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    6562..6595
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        6612..6658
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
ORIGIN
        1 tccggtattt ttacaacaat taccacaaca aaacaaacaa caaacaacat tacaatttac
       61 tattctagtc gaatactgta tttttacaac aattaccaac aacaacaaac aacaaacaac
      121 attacaatta ctatttacaa ttacaatgga gatggatatg gatatcatta gaaaggcaat
      181 tgctgccact atctttgcat tggttatggc ttgggcgtgg agagtgttgg attgggcatg
      241 gtttactcct aagaggatcg agaaacgtct aaggcagcaa ggttttagag gaaatcctta
      301 tagattcttg gttggagatg ttaaggagag tggaaaaatg catcaagaag ccttgtctaa
      361 ccccatggag ttcgacaacg atattgttcc tcgcctcatg ccacatatta accacaccat
      421 aaaaacttac ggtaggaatt catttacatg gatgggaagg attccaagga ttcatgttat
      481 ggaacctgaa cttattaagg aagtattgac ccactccagc aaataccaaa agaactttga
      541 tgttcacaat ccccttgtta agttccttct cactggagtt ggaagctttg aaggggcaaa
      601 atggtccaaa cacagaagaa tcatttcccc tgccttcact cttgagaaac taaagtcaat
      661 gttgccggct tttgccatat gctatcatga catgttgacg aaatgggaga aattagcaga
      721 aaaagaagga tcccatgaag ttgatatctt ccccacattt gatgttttga caagtgatgt
      781 aatttcaaag gttgcatttg gtagcacata cgatgaagga ggcaaaatct tcagactatt
      841 gaaagaactc atggatctca caattgattg catgagagat gtctacatcc cagggtggag
      901 ctacttgcca accaagagga acaagaggat gaaagaaatt aacaaagaaa tcacagatat
      961 gctaaggttc atcatcaaca agagaatgaa ggctttgaag gctggagaac caggtgagga
     1021 tgacttgttg ggagtcctgt tggaatcaaa cattcaagaa attcagaaac aaggaaacag
     1081 gaaggatggt ggaatgacaa tcaacgatgt gattgaggaa tgcaaattgt tctactttgc
     1141 tggtcaagaa accaccggag ttttgctcac atggaccacc attttattga gcaagcatcc
     1201 cgagtggcaa gagcgagcta gggaagaagt tctccaagcc tttggcaaga acaagcctga
     1261 atttgaacgc ttgaatcatc ttaaatatgt ttctatgatc ttatacgagg ttctaaggtt
     1321 gtacccgcca gttattgatc tgaccaagat tattcacgag gacacaaagt tagggccgta
     1381 cacaatccct gcaggaacac aagtaatgtt gccgacagta atgcttcata gagagaagtc
     1441 catttgggga gaagatgcaa tggaattcaa cccaatgaga tttgctgatg gagttgccaa
     1501 tgccaccaag aacaatgtca catatttgcc attcagttgg ggacctagag tttgcctcgg
     1561 ccaaaacttt gcacttcttc aagcaaaatt agggttggca atgattttac aacgcttcaa
     1621 atttgatgta gctccatcct atgttcatgc tccctttacc attcttacag ttcaacctca
     1681 gtttggttct catgttatct acaagaagct tgagagacag aatttttaag cttgtcaagc
     1741 agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     1801 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     1861 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     1921 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     1981 tatgttacta gatcgacgct tgtgttgtct tctgcacgaa gtggtttaaa ctatcagtgt
     2041 ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga
     2101 tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac
     2161 agggttcccc agatcaggcg ctggctgctg aacccccagc cggaactgac cccacaaggc
     2221 cctagcgttt gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc
     2281 ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat
     2341 ccgatccgca catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc
     2401 tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga
     2461 atttcgtgta gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc
     2521 aacgggacgt tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt
     2581 ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc
     2641 attcctcggc cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct
     2701 cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct
     2761 gctgccacac cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca
     2821 cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt
     2881 tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg
     2941 gccacggcgc aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt
     3001 tcagcaacgc ggcctgcttg gcctcgctga cctgttttgc caggtcctcg ccggcggttt
     3061 ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg
     3121 ccgcctcctg ttcaagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag
     3181 ggggagccag ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg accatcgagc
     3241 cgacggactg gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg
     3301 catcctcggc ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg
     3361 tccgattcat tcaccctcct tgcgggattg ccccgactca cgccggggca atgtgccctt
     3421 attcctgatt tgacccgcct ggtgccttgg tgtccagata atccacctta tcggcaatga
     3481 agtcggtccc gtagaccgtc tggccgtcct tctcgtactt ggtattccga atcttgccct
     3541 gcacgaatac cagcgacccc ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa
     3601 aacacttgat gcggaagaag tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca
     3661 tctaggatct gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3721 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3781 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3841 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3901 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3961 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4021 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4081 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4141 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4201 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4261 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4321 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4381 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4441 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4501 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4561 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagaac cacgctcacc
     4621 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4681 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4741 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4801 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4861 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4921 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4981 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5041 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5101 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5161 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5221 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5281 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5341 atattattga agcatttatc aaggttattg tctcatgagc ggatacatat ttgaatgtat
     5401 ttagaaaaat aaacaaatag gggttccgcg cacgaattgg ccagcgctgc catttttggg
     5461 gtgaggccgt tcgcggccga ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg
     5521 ggccgggagg gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag
     5581 ggcgcagccc tggttaaaaa caaggtttat aaatattggt ttaaaagcag gttaaaagac
     5641 aggttagcgg tggccgaaaa acgggcggaa acccttgcaa atgctggatt ttctgcctgt
     5701 ggacagcccc tcaaatgtca ataggtgcgc ccctcatctg tcagcactct gcccctcaag
     5761 tgtcaaggat cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag
     5821 ggcacttatc cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt
     5881 ttcgccgatt tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag cctgcccctc
     5941 atctgtcaac gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct
     6001 gtcagtgagg gccaagtttt ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc
     6061 acacggcttc gacggcgttt ctggcgcgtt tgcagggcca tagacggccg ccagcccagc
     6121 ggcgagggca accagcccgg tgagcgtcgc aaaggagatc ctgatctgac tgatgggctg
     6181 cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt
     6241 ggcaggatat attgtggtgt aaacaaattg acgcttagac aacttaataa cacattgcgg
     6301 acgtttttaa tgtactgggg tggatgcagt gggccccact ctgtgaagac aacagagaat
     6361 tcgcatgcgg agcggtgcgg gtgccagggc gtgcccttgg gctccccggg cgcgtactcc
     6421 actccccgag ccggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca
     6481 ctccccccat ggtgcggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact
     6541 ccactccccg agccggtgcg ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc
     6601 cactcccctt cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggac
//
