LOCUS       sequence-349451-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 6, transcriptional unit for transient 
            expression of 7-deoxyloganetic acid glucosyl transferase (Cr7-DLGT) 
            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 6524)
  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 6524)
  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: pEPQD1CB0861
FEATURES             Location/Qualifiers
     source          1..6524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    112..136
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    253..286
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    312..345
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    375..408
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    434..467
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        484..530
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    620..675
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      2138..2390
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2438..2462
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2618..3766)
                     /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(4102..4690)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4182..4201)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4861..5652,5653..5721))
                     /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     5484..5503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5722..5826)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(5913..6524,1..20)
                     /label=oriV
                     /note="incP origin of replication"
ORIGIN
        1 caaccagccc ggtgagcgtc gcaaaggaga tcctgatctg actgatgggc tgcctgtatc
       61 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat
      121 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt
      181 aatgtactgg ggtggatgca gtgggcccca ctctgtgaag acaatgtgga attcctcgag
      241 ggagcggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact ccactccccg
      301 agccggtgcg ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc cactcccccc
      361 atggtgcggt gcgggtgcca gggcgtgccc ttgggctccc cgggcgcgta ctccactccc
      421 cgagccggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac tccactcccc
      481 ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga ctccggtatt
      541 tttacaacaa ttaccacaac aaaacaaaca acaaacaaca ttacaattta ctattctagt
      601 cgaatactgt atttttacaa caattaccaa caacaacaaa caacaaacaa cattacaatt
      661 actatttaca attacaatgg gttctcaaga aacaaatttg gctcctcatg ttctcatatt
      721 ccctttgcca attcaaggcc atgtgaactc catgctcaga ctagcagaac ttctctgtct
      781 agctgaatta gatatcactt ttattgtttc agagttcagc cacagtcgct taattaagca
      841 tacaaatgtt gcctcccgtt ttgcccgtta cccaggattt cagttccagc ccatatccga
      901 tggccttccc gatgatcacc ccagagcagg cgaaagagtc atggatattt tgccttcaac
      961 taaaaatgta acgggacccc tgtttaagca gatgatggtt gagaataagt gcttttcttc
     1021 tgctactcga aggccgatta cttgcataat cgctgatggg gttttgagct ttgctgggga
     1081 tttcgctcaa gaaaaaggaa tccctcttat ttacttcaga actgtttctg cttgttcttt
     1141 ctgggcttgt ttttgtatgc ctgaactaat tgaatctggt gacattccga ttaaaggaaa
     1201 tggtatggat ctgatagtga aaagcgtgcc ggggatggaa acttttctca ggcgacgaga
     1261 tctgccgggg ttttgccgag tcaatgacat taatgaaccg aaactccaga tcctgaaaac
     1321 agagacaaga cagacaacga gagcccaagc ggctattttg aacacttttg aggatttgga
     1381 aggaccaatt ctatctcaaa ttcgcaaaca catgccaaga cttttcacaa ttggaccaag
     1441 tcattcccac ctaacatcta gacttgaaac caagaatatc aagactttaa tttcctccgg
     1501 tagtttctgg gaagaagatc gaagctgtgt ggattggctt gatgcacagc caccaagatc
     1561 tgttttatat gtaagttttg gaagtattac agttgtaaca agagatcaac tcctggaatt
     1621 ctggtatggt cttgttaata gtggacaacg gttcttgtgg gtaatgaggc cagattcaat
     1681 tatgggcaaa gatgggcaaa gccagattcc tgcagatctt gaagagggca caaaagccag
     1741 aggttatatg gtgggatggg cacctcaaga agaggtcttg aatcaccctg ccattggagg
     1801 atttctgact cacagtgggt ggaattcaac tttagagagt attgtggctg gtgtcccaat
     1861 gatatgttgg ccatactttg cggaccaaat gataaacagc agatttgtga gtgaaatttg
     1921 gaagattgga ttggatatga aagatacatg tgatagagag acaattgtga agatggttag
     1981 ggagttgatg gagattagga aagatgagtt cttgcaaagg gctgatcata tggctaaatt
     2041 ggcaaaagag gctgttagtg aaggcggatc ctcatattct aacttggatg gcctcgttga
     2101 ttacataaaa tcactgatta tttgagcttg tcaagcagat cgttcaaaca tttggcaata
     2161 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
     2221 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
     2281 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
     2341 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gacgctgagc
     2401 ttgtcttctg cacgaagtgg tttaaactat cagtgtttga caggatatat tggcgggtaa
     2461 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
     2521 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttccccagat caggcgctgg
     2581 ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa tgcaccaggt
     2641 catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg caggcttcgc
     2701 cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg aggcggaagg
     2761 tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac atccgtcggg
     2821 ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg tcgccagcaa
     2881 acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc ttgccacggt
     2941 ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg cggtcggacg
     3001 tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc gtgtaatacc
     3061 ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag gtgatcggct
     3121 cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt tcgtcatcgt
     3181 cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg tggaaaatga
     3241 ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac agggcagagc
     3301 gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata tcgaacaagg
     3361 aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc tgcttggcct
     3421 cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc gtcatagttc
     3481 ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttca agacgacgcg
     3541 aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc acgctgtcgc
     3601 gctcgatctt ggccgtagct tgctggacca tcgagccgac ggactggaag gtttcgcggg
     3661 gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa aaccccgcgt
     3721 cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac cctccttgcg
     3781 ggattgcccc gactcacgcc ggggcaatgt gcccttattc ctgatttgac ccgcctggtg
     3841 ccttggtgtc cagataatcc accttatcgg caatgaagtc ggtcccgtag accgtctggc
     3901 cgtccttctc gtacttggta ttccgaatct tgccctgcac gaataccagc gaccccttgc
     3961 ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg aagaagtcgg
     4021 tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggatctgcca ggaaccgtaa
     4081 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     4141 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4201 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4261 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4321 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4381 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4441 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4501 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4561 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4621 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4681 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4741 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4801 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4861 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4921 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4981 cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa
     5041 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     5101 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     5161 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5221 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5281 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5341 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5401 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5461 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5521 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5581 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5641 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5701 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaagg
     5761 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5821 tccgcgcacg aattggccag cgctgccatt tttggggtga ggccgttcgc ggccgagggg
     5881 cgcagcccct ggggggatgg gaggcccgcg ttagcgggcc gggagggttc gagaaggggg
     5941 ggcacccccc ttcggcgtgc gcggtcacgc gcacagggcg cagccctggt taaaaacaag
     6001 gtttataaat attggtttaa aagcaggtta aaagacaggt tagcggtggc cgaaaaacgg
     6061 gcggaaaccc ttgcaaatgc tggattttct gcctgtggac agcccctcaa atgtcaatag
     6121 gtgcgcccct catctgtcag cactctgccc ctcaagtgtc aaggatcgcg cccctcatct
     6181 gtcagtagtc gcgcccctca agtgtcaata ccgcagggca cttatcccca ggcttgtcca
     6241 catcatctgt gggaaactcg cgtaaaatca ggcgttttcg ccgatttgcg aggctggcca
     6301 gctccacgtc gccggccgaa atcgagcctg cccctcatct gtcaacgccg cgccgggtga
     6361 gtcggcccct caagtgtcaa cgtccgcccc tcatctgtca gtgagggcca agttttccgc
     6421 gaggtatcca caacgccggc ggccgcggtg tctcgcacac ggcttcgacg gcgtttctgg
     6481 cgcgtttgca gggccataga cggccgccag cccagcggcg aggg
//
