LOCUS       sequence-349546-        6524 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 3, 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: pEPQD1CB0867
FEATURES             Location/Qualifiers
     source          1..6524
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     290..309
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(528..632)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      719..1350
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    1442..1466
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    1583..1616
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    1642..1675
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    1705..1738
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    1764..1797
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        1814..1860
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    1950..2005
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      3468..3720
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3768..3792
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(3948..5096)
                     /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(5432..6020)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5512..5531)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6191..6524,1..458,459..527))
                     /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"
ORIGIN
        1 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
       61 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
      121 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
      181 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
      241 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
      301 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
      361 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
      421 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
      481 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
      541 caatattatt gaagcattta tcaaggttat tgtctcatga gcggatacat atttgaatgt
      601 atttagaaaa ataaacaaat aggggttccg cgcacgaatt ggccagcgct gccatttttg
      661 gggtgaggcc gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag
      721 cgggccggga gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac
      781 agggcgcagc cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag
      841 acaggttagc ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct
      901 gtggacagcc cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca
      961 agtgtcaagg atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc
     1021 agggcactta tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg
     1081 ttttcgccga tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc
     1141 tcatctgtca acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat
     1201 ctgtcagtga gggccaagtt ttccgcgagg tatccacaac gccggcggcc gcggtgtctc
     1261 gcacacggct tcgacggcgt ttctggcgcg tttgcagggc catagacggc cgccagccca
     1321 gcggcgaggg caaccagccc ggtgagcgtc gcaaaggaga tcctgatctg actgatgggc
     1381 tgcctgtatc gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg
     1441 gtggcaggat atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc
     1501 ggacgttttt aatgtactgg ggtggatgca gtgggcccca ctctgtgaag acaaactaga
     1561 attcgagctc ggagcggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact
     1621 ccactccccg agccggtgcg ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc
     1681 cactcccccc atggtgcggt gcgggtgcca gggcgtgccc ttgggctccc cgggcgcgta
     1741 ctccactccc cgagccggtg cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac
     1801 tccactcccc ttcgcaagac ccttcctcta tataaggaag ttcatttcat ttggagagga
     1861 ctccggtatt tttacaacaa ttaccacaac aaaacaaaca acaaacaaca ttacaattta
     1921 ctattctagt cgaatactgt atttttacaa caattaccaa caacaacaaa caacaaacaa
     1981 cattacaatt actatttaca attacaatgg gttctcaaga aacaaatttg gctcctcatg
     2041 ttctcatatt ccctttgcca attcaaggcc atgtgaactc catgctcaga ctagcagaac
     2101 ttctctgtct agctgaatta gatatcactt ttattgtttc agagttcagc cacagtcgct
     2161 taattaagca tacaaatgtt gcctcccgtt ttgcccgtta cccaggattt cagttccagc
     2221 ccatatccga tggccttccc gatgatcacc ccagagcagg cgaaagagtc atggatattt
     2281 tgccttcaac taaaaatgta acgggacccc tgtttaagca gatgatggtt gagaataagt
     2341 gcttttcttc tgctactcga aggccgatta cttgcataat cgctgatggg gttttgagct
     2401 ttgctgggga tttcgctcaa gaaaaaggaa tccctcttat ttacttcaga actgtttctg
     2461 cttgttcttt ctgggcttgt ttttgtatgc ctgaactaat tgaatctggt gacattccga
     2521 ttaaaggaaa tggtatggat ctgatagtga aaagcgtgcc ggggatggaa acttttctca
     2581 ggcgacgaga tctgccgggg ttttgccgag tcaatgacat taatgaaccg aaactccaga
     2641 tcctgaaaac agagacaaga cagacaacga gagcccaagc ggctattttg aacacttttg
     2701 aggatttgga aggaccaatt ctatctcaaa ttcgcaaaca catgccaaga cttttcacaa
     2761 ttggaccaag tcattcccac ctaacatcta gacttgaaac caagaatatc aagactttaa
     2821 tttcctccgg tagtttctgg gaagaagatc gaagctgtgt ggattggctt gatgcacagc
     2881 caccaagatc tgttttatat gtaagttttg gaagtattac agttgtaaca agagatcaac
     2941 tcctggaatt ctggtatggt cttgttaata gtggacaacg gttcttgtgg gtaatgaggc
     3001 cagattcaat tatgggcaaa gatgggcaaa gccagattcc tgcagatctt gaagagggca
     3061 caaaagccag aggttatatg gtgggatggg cacctcaaga agaggtcttg aatcaccctg
     3121 ccattggagg atttctgact cacagtgggt ggaattcaac tttagagagt attgtggctg
     3181 gtgtcccaat gatatgttgg ccatactttg cggaccaaat gataaacagc agatttgtga
     3241 gtgaaatttg gaagattgga ttggatatga aagatacatg tgatagagag acaattgtga
     3301 agatggttag ggagttgatg gagattagga aagatgagtt cttgcaaagg gctgatcata
     3361 tggctaaatt ggcaaaagag gctgttagtg aaggcggatc ctcatattct aacttggatg
     3421 gcctcgttga ttacataaaa tcactgatta tttgagcttg tcaagcagat cgttcaaaca
     3481 tttggcaata aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat
     3541 aatttctgtt gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta
     3601 tgagatgggt ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca
     3661 aaatatagcg cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc
     3721 gacgctttac ttgtcttctg cacgaagtgg tttaaactat cagtgtttga caggatatat
     3781 tggcgggtaa acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg
     3841 tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttccccagat
     3901 caggcgctgg ctgctgaacc cccagccgga actgacccca caaggcccta gcgtttgcaa
     3961 tgcaccaggt catcattgac ccaggcgtgt tccaccaggc cgctgcctcg caactcttcg
     4021 caggcttcgc cgacctgctc gcgccacttc ttcacgcggg tggaatccga tccgcacatg
     4081 aggcggaagg tttccagctt gagcgggtac ggctcccggt gcgagctgaa atagtcgaac
     4141 atccgtcggg ccgtcggcga cagcttgcgg tacttctccc atatgaattt cgtgtagtgg
     4201 tcgccagcaa acagcacgac gatttcctcg tcgatcagga cctggcaacg ggacgttttc
     4261 ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca ccgattccag gtgcccaacg
     4321 cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg acaggcattc ctcggccttc
     4381 gtgtaatacc ggccattgat cgaccagccc aggtcctggc aaagctcgta gaacgtgaag
     4441 gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca acacctgctg ccacaccagt
     4501 tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga tcttcacgtc cttgttgacg
     4561 tggaaaatga ccttgttttg cagcgcctcg cgcgggattt tcttgttgcg cgtggtgaac
     4621 agggcagagc gggccgtgtc gtttggcatc gctcgcatcg tgtccggcca cggcgcaata
     4681 tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg tgtgtttcag caacgcggcc
     4741 tgcttggcct cgctgacctg ttttgccagg tcctcgccgg cggtttttcg cttcttggtc
     4801 gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca aacctgccgc ctcctgttca
     4861 agacgacgcg aacgctccac ggcggccgat ggcgcgggca gggcaggggg agccagttgc
     4921 acgctgtcgc gctcgatctt ggccgtagct tgctggacca tcgagccgac ggactggaag
     4981 gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg tttcggcatc ctcggcggaa
     5041 aaccccgcgt cgatcagttc ttgcctgtat gccttccggt caaacgtccg attcattcac
     5101 cctccttgcg ggattgcccc gactcacgcc ggggcaatgt gcccttattc ctgatttgac
     5161 ccgcctggtg ccttggtgtc cagataatcc accttatcgg caatgaagtc ggtcccgtag
     5221 accgtctggc cgtccttctc gtacttggta ttccgaatct tgccctgcac gaataccagc
     5281 gaccccttgc ccaaatactt gccgtgggcc tcggcctgag agccaaaaca cttgatgcgg
     5341 aagaagtcgg tgcgctcctg cttgtcgccg gcatcgttgc gccacatcta ggatctgcca
     5401 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5461 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5521 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5581 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5641 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5701 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5761 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5821 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5881 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5941 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6001 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6061 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6121 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6181 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6241 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6301 catctggccc cagtgctgca atgataccgc gagaaccacg ctcaccggct ccagatttat
     6361 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6421 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6481 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtca
//
