LOCUS       sequence-349439-        6659 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 2, 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: pEPQD1CB0871
FEATURES             Location/Qualifiers
     source          1..6659
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    1..25
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    142..175
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    201..234
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    264..297
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    323..356
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        373..419
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    509..564
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      2162..2414
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2462..2486
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2642..3790)
                     /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(4126..4714)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4206..4225)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4885..5676,5677..5745))
                     /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     5508..5527
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5746..5850)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5937..6568
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 tggcaggata tattgtggtg taaacaaatt gacgcttaga caacttaata acacattgcg
       61 gacgttttta atgtactggg gtggatgcag tgggccccac tctgtgaaga caagcaagaa
      121 ttcaagcttg gagcggtgcg ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc
      181 cactccccga gccggtgcgg gtgccagggc gtgcccttgg gctccccggg cgcgtactcc
      241 actcccccca tggtgcggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac
      301 tccactcccc gagccggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact
      361 ccactcccct tcgcaagacc cttcctctat ataaggaagt tcatttcatt tggagaggac
      421 tccggtattt ttacaacaat taccacaaca aaacaaacaa caaacaacat tacaatttac
      481 tattctagtc gaatactgta tttttacaac aattaccaac aacaacaaac aacaaacaac
      541 attacaatta ctatttacaa ttacaatgga gatggatatg gatatcatta gaaaggcaat
      601 tgctgccact atctttgcat tggttatggc ttgggcgtgg agagtgttgg attgggcatg
      661 gtttactcct aagaggatcg agaaacgtct aaggcagcaa ggttttagag gaaatcctta
      721 tagattcttg gttggagatg ttaaggagag tggaaaaatg catcaagaag ccttgtctaa
      781 ccccatggag ttcgacaacg atattgttcc tcgcctcatg ccacatatta accacaccat
      841 aaaaacttac ggtaggaatt catttacatg gatgggaagg attccaagga ttcatgttat
      901 ggaacctgaa cttattaagg aagtattgac ccactccagc aaataccaaa agaactttga
      961 tgttcacaat ccccttgtta agttccttct cactggagtt ggaagctttg aaggggcaaa
     1021 atggtccaaa cacagaagaa tcatttcccc tgccttcact cttgagaaac taaagtcaat
     1081 gttgccggct tttgccatat gctatcatga catgttgacg aaatgggaga aattagcaga
     1141 aaaagaagga tcccatgaag ttgatatctt ccccacattt gatgttttga caagtgatgt
     1201 aatttcaaag gttgcatttg gtagcacata cgatgaagga ggcaaaatct tcagactatt
     1261 gaaagaactc atggatctca caattgattg catgagagat gtctacatcc cagggtggag
     1321 ctacttgcca accaagagga acaagaggat gaaagaaatt aacaaagaaa tcacagatat
     1381 gctaaggttc atcatcaaca agagaatgaa ggctttgaag gctggagaac caggtgagga
     1441 tgacttgttg ggagtcctgt tggaatcaaa cattcaagaa attcagaaac aaggaaacag
     1501 gaaggatggt ggaatgacaa tcaacgatgt gattgaggaa tgcaaattgt tctactttgc
     1561 tggtcaagaa accaccggag ttttgctcac atggaccacc attttattga gcaagcatcc
     1621 cgagtggcaa gagcgagcta gggaagaagt tctccaagcc tttggcaaga acaagcctga
     1681 atttgaacgc ttgaatcatc ttaaatatgt ttctatgatc ttatacgagg ttctaaggtt
     1741 gtacccgcca gttattgatc tgaccaagat tattcacgag gacacaaagt tagggccgta
     1801 cacaatccct gcaggaacac aagtaatgtt gccgacagta atgcttcata gagagaagtc
     1861 catttgggga gaagatgcaa tggaattcaa cccaatgaga tttgctgatg gagttgccaa
     1921 tgccaccaag aacaatgtca catatttgcc attcagttgg ggacctagag tttgcctcgg
     1981 ccaaaacttt gcacttcttc aagcaaaatt agggttggca atgattttac aacgcttcaa
     2041 atttgatgta gctccatcct atgttcatgc tccctttacc attcttacag ttcaacctca
     2101 gtttggttct catgttatct acaagaagct tgagagacag aatttttaag cttgtcaagc
     2161 agatcgttca aacatttggc aataaagttt cttaagattg aatcctgttg ccggtcttgc
     2221 gatgattatc atataatttc tgttgaatta cgttaagcat gtaataatta acatgtaatg
     2281 catgacgtta tttatgagat gggtttttat gattagagtc ccgcaattat acatttaata
     2341 cgcgatagaa aacaaaatat agcgcgcaaa ctaggataaa ttatcgcgcg cggtgtcatc
     2401 tatgttacta gatcgacgct actattgtct tctgcacgaa gtggtttaaa ctatcagtgt
     2461 ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga
     2521 tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac
     2581 agggttcccc agatcaggcg ctggctgctg aacccccagc cggaactgac cccacaaggc
     2641 cctagcgttt gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc
     2701 ctcgcaactc ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat
     2761 ccgatccgca catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc
     2821 tgaaatagtc gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga
     2881 atttcgtgta gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc
     2941 aacgggacgt tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt
     3001 ccaggtgccc aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc
     3061 attcctcggc cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct
     3121 cgtagaacgt gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct
     3181 gctgccacac cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca
     3241 cgtccttgtt gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt
     3301 tgcgcgtggt gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg
     3361 gccacggcgc aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt
     3421 tcagcaacgc ggcctgcttg gcctcgctga cctgttttgc caggtcctcg ccggcggttt
     3481 ttcgcttctt ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg
     3541 ccgcctcctg ttcaagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag
     3601 ggggagccag ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg accatcgagc
     3661 cgacggactg gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg
     3721 catcctcggc ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg
     3781 tccgattcat tcaccctcct tgcgggattg ccccgactca cgccggggca atgtgccctt
     3841 attcctgatt tgacccgcct ggtgccttgg tgtccagata atccacctta tcggcaatga
     3901 agtcggtccc gtagaccgtc tggccgtcct tctcgtactt ggtattccga atcttgccct
     3961 gcacgaatac cagcgacccc ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa
     4021 aacacttgat gcggaagaag tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca
     4081 tctaggatct gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4141 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4201 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4261 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4321 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4381 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4441 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4501 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4561 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4621 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4681 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4741 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4801 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4861 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4921 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4981 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagaac cacgctcacc
     5041 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     5101 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     5161 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     5221 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     5281 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     5341 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     5401 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     5461 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     5521 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5581 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5641 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5701 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5761 atattattga agcatttatc aaggttattg tctcatgagc ggatacatat ttgaatgtat
     5821 ttagaaaaat aaacaaatag gggttccgcg cacgaattgg ccagcgctgc catttttggg
     5881 gtgaggccgt tcgcggccga ggggcgcagc ccctgggggg atgggaggcc cgcgttagcg
     5941 ggccgggagg gttcgagaag ggggggcacc ccccttcggc gtgcgcggtc acgcgcacag
     6001 ggcgcagccc tggttaaaaa caaggtttat aaatattggt ttaaaagcag gttaaaagac
     6061 aggttagcgg tggccgaaaa acgggcggaa acccttgcaa atgctggatt ttctgcctgt
     6121 ggacagcccc tcaaatgtca ataggtgcgc ccctcatctg tcagcactct gcccctcaag
     6181 tgtcaaggat cgcgcccctc atctgtcagt agtcgcgccc ctcaagtgtc aataccgcag
     6241 ggcacttatc cccaggcttg tccacatcat ctgtgggaaa ctcgcgtaaa atcaggcgtt
     6301 ttcgccgatt tgcgaggctg gccagctcca cgtcgccggc cgaaatcgag cctgcccctc
     6361 atctgtcaac gccgcgccgg gtgagtcggc ccctcaagtg tcaacgtccg cccctcatct
     6421 gtcagtgagg gccaagtttt ccgcgaggta tccacaacgc cggcggccgc ggtgtctcgc
     6481 acacggcttc gacggcgttt ctggcgcgtt tgcagggcca tagacggccg ccagcccagc
     6541 ggcgagggca accagcccgg tgagcgtcgc aaaggagatc ctgatctgac tgatgggctg
     6601 cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctgg
//
