LOCUS       sequence-349532-        7294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 2, transcriptional unit for transient 
            expression of 1-deoxy-D-xylulose 5-phosphate synthase (CrDXS2) 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 7294)
  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 7294)
  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: pEPQD1CB0785
FEATURES             Location/Qualifiers
     source          1..7294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(93..884,885..953))
                     /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     716..735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(954..1058)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1145..1776
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    1868..1892
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    2009..2042
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    2068..2101
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    2131..2164
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    2190..2223
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        2240..2286
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     terminator      4666..4918
                     /label=MAS terminator
                     /note="mannopine synthase terminator"
     misc_feature    4964..4988
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(5144..6292)
                     /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(6628..7216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6708..6727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
       61 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      121 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
      181 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaacca
      241 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      301 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      361 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      421 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      481 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
      541 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
      601 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
      661 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
      721 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
      781 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
      841 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
      901 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
      961 ctttttcaat attattgaag catttatcaa ggttattgtc tcatgagcgg atacatattt
     1021 gaatgtattt agaaaaataa acaaataggg gttccgcgca cgaattggcc agcgctgcca
     1081 tttttggggt gaggccgttc gcggccgagg ggcgcagccc ctggggggat gggaggcccg
     1141 cgttagcggg ccgggagggt tcgagaaggg ggggcacccc ccttcggcgt gcgcggtcac
     1201 gcgcacaggg cgcagccctg gttaaaaaca aggtttataa atattggttt aaaagcaggt
     1261 taaaagacag gttagcggtg gccgaaaaac gggcggaaac ccttgcaaat gctggatttt
     1321 ctgcctgtgg acagcccctc aaatgtcaat aggtgcgccc ctcatctgtc agcactctgc
     1381 ccctcaagtg tcaaggatcg cgcccctcat ctgtcagtag tcgcgcccct caagtgtcaa
     1441 taccgcaggg cacttatccc caggcttgtc cacatcatct gtgggaaact cgcgtaaaat
     1501 caggcgtttt cgccgatttg cgaggctggc cagctccacg tcgccggccg aaatcgagcc
     1561 tgcccctcat ctgtcaacgc cgcgccgggt gagtcggccc ctcaagtgtc aacgtccgcc
     1621 cctcatctgt cagtgagggc caagttttcc gcgaggtatc cacaacgccg gcggccgcgg
     1681 tgtctcgcac acggcttcga cggcgtttct ggcgcgtttg cagggccata gacggccgcc
     1741 agcccagcgg cgagggcaac cagcccggtg agcgtcgcaa aggagatcct gatctgactg
     1801 atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc
     1861 tggctggtgg caggatatat tgtggtgtaa acaaattgac gcttagacaa cttaataaca
     1921 cattgcggac gtttttaatg tactggggtg gatgcagtgg gccccactct gtgaagacaa
     1981 gcaagaattc aagcttggag cggtgcgggt gccagggcgt gcccttgggc tccccgggcg
     2041 cgtactccac tccccgagcc ggtgcgggtg ccagggcgtg cccttgggct ccccgggcgc
     2101 gtactccact ccccccatgg tgcggtgcgg gtgccagggc gtgcccttgg gctccccggg
     2161 cgcgtactcc actccccgag ccggtgcggg tgccagggcg tgcccttggg ctccccgggc
     2221 gcgtactcca ctccccttcg caagaccctt cctctatata aggaagttca tttcatttgg
     2281 agaggactcc ggtattttta caacaattac cacaacaaaa caaacaacaa acaacattac
     2341 aatttactat tctagtcgaa tactgtataa gagctcattt ttacaacaat taccaacaac
     2401 aacaaacaac aaacaacatt acaattacat ttacaattat cgataacatg gcttcttcta
     2461 tgctttcttc tgctgctgtt gttgctactc gtgctagtgc tgctcaagct agtatggttg
     2521 ctccttttac tggacttaag tctgctgctt cttttcctgt tactagaaag caaaacaacc
     2581 ttgatattac ttctattgct agtaacggag gaagagttcg agcaatggct gctgctcatg
     2641 ggcatgctca taatgttgat gaaaattacc aagtcctagt gaagaagcag cctttgaatt
     2701 tttcaggaga taagcctttt actcctattt tggataccat caactaccct acccatatga
     2761 agaatctctc tcttcaggag cttcaacaat tagctgacga attaagggag gagatagtgt
     2821 attctgtggc caaaacgggt ggtcacttaa gttccagcct aggtgtggcc gagctcactg
     2881 tggcactcca ccacgtgttc gaaaccccaa atgataaaat catatgggat gttggccacc
     2941 agacatatgg gcataagatt ctgacaggga gaaggtcaaa gatgcatacc atgagacaaa
     3001 cttgcggact tgcaggtttc cccaaaagag atgaaagtgc ccatgatgca tttggagttg
     3061 gccatagttc caccagcatt tctgccggtt taggaatggc aataggacga gatttattag
     3121 gtaagaaaaa tcatgtaata acggtaatag gagatggagc tatgacagcc ggacaagcat
     3181 atgaagccat gaataatgct ggatatcttg attctaatct cataatcata ttgaatgata
     3241 acaaacaagt ttctctcccg actgccacaa ttgatggtcc tgcacctcct actggtgctc
     3301 taagcaatgc cttaactagg cttcaatcta gtagaaaatt tcgactactt cgggaggctg
     3361 caaaaggtgt gacaaagcaa ttagggggac aagttcatga aatggcagct aaagtggata
     3421 catatatgag agggatgaat agccaaaggg cttccttatt tgaagattta ggaatgtatt
     3481 atattggtcc acttgatggt cacaatatgg aagatcttgt tgatatttta gagaaagtaa
     3541 aatctgttcc tgcaacgggc cctgttctta ttcatattat tactgagaaa ggaaaaggat
     3601 atcctcctgc tgaagctgcc gccgacaaaa tgcatggggt ggtgaaattt gagccaatga
     3661 caggaaaaca attcaagtca aagtcaaaaa caaagtcata tacaaattat tttgctgaga
     3721 gtttaataag tgaagctagg gaagatgata agattattgc cattcatgct gctatgggag
     3781 gaggaacagg actcaattta ttccagaaac actttcctga acgatgcttt gatgttggga
     3841 tagctgagca acatgctgtc acttttgctg ctggcttagc ttctgaagga ctcaagcctt
     3901 tttgtgctat ttactcatct ttcttacaaa gaggttatga tcaggtggtt catgatgtgg
     3961 accttcaaaa aattccagta aggtttgcta tggatagagc agggctggtg ggtgcagatg
     4021 gacccactca ttgcggtgca tttgacacaa cttttatggc ttgtttgcct aatatggtgg
     4081 tcatggctcc ttcagatgaa accgaattga tgcacatggt tgccacagca gctgccattg
     4141 atgataggcc tagttgcttt cgatatccca gaggaaatgg cattggtgca attcttccac
     4201 caaacaacaa aggaactcca ttacagattg gtaaaggaag aatattgttg gagggaacaa
     4261 gagttgccat tcttgggtat ggagcaatgg tgcaaaactg tttaatggct gctcagcttc
     4321 ttgaaattaa tggaatatca gctacagttg ttgatgcccg attttgcaaa cctcttgatg
     4381 gagatttgat tagaaaattg gctcaacaac atgaagtcct tatcactgtt gaagaaggat
     4441 ccattggtgg ttttggttcc catgtggcac agttcttggc tttgaatgga ttgcttgatg
     4501 gaaatttaaa gtggagggca atgatgcttc ctgacagata tattgatcat ggagctcaaa
     4561 atgaccaaat tgaagaggct ggcctaactc caaagcatat tgctggcact gtgttttctc
     4621 tactcagcaa aaacaagaat actcttcaag ttctccagtt ttaggcttgg actcccatgt
     4681 tggcaaaggc aaccaaacaa acaatgaatg atccgctcct gcatatgggg cggtttgagt
     4741 atttcaactg ccatttgggc tgaattgtag acatgctcct gtcagaaatt ccgtgatctt
     4801 actcaatatt cagtaatctc ggccaatatc ctaaatgtgc gtggctttat ctgtctttgt
     4861 attgtttcat caattcatgt aacgtttgct tttcttatga attttcaaat aaattatccg
     4921 ctactattgt cttctgcacg aagtggttta aactatcagt gtttgacagg atatattggc
     4981 gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa agggcgtgaa
     5041 aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc ccagatcagg
     5101 cgctggctgc tgaaccccca gccggaactg accccacaag gccctagcgt ttgcaatgca
     5161 ccaggtcatc attgacccag gcgtgttcca ccaggccgct gcctcgcaac tcttcgcagg
     5221 cttcgccgac ctgctcgcgc cacttcttca cgcgggtgga atccgatccg cacatgaggc
     5281 ggaaggtttc cagcttgagc gggtacggct cccggtgcga gctgaaatag tcgaacatcc
     5341 gtcgggccgt cggcgacagc ttgcggtact tctcccatat gaatttcgtg tagtggtcgc
     5401 cagcaaacag cacgacgatt tcctcgtcga tcaggacctg gcaacgggac gttttcttgc
     5461 cacggtccag gacgcggaag cggtgcagca gcgacaccga ttccaggtgc ccaacgcggt
     5521 cggacgtgaa gcccatcgcc gtcgcctgta ggcgcgacag gcattcctcg gccttcgtgt
     5581 aataccggcc attgatcgac cagcccaggt cctggcaaag ctcgtagaac gtgaaggtga
     5641 tcggctcgcc gataggggtg cgcttcgcgt actccaacac ctgctgccac accagttcgt
     5701 catcgtcggc ccgcagctcg acgccggtgt aggtgatctt cacgtccttg ttgacgtgga
     5761 aaatgacctt gttttgcagc gcctcgcgcg ggattttctt gttgcgcgtg gtgaacaggg
     5821 cagagcgggc cgtgtcgttt ggcatcgctc gcatcgtgtc cggccacggc gcaatatcga
     5881 acaaggaaag ctgcatttcc ttgatctgct gcttcgtgtg tttcagcaac gcggcctgct
     5941 tggcctcgct gacctgtttt gccaggtcct cgccggcggt ttttcgcttc ttggtcgtca
     6001 tagttcctcg cgtgtcgatg gtcatcgact tcgccaaacc tgccgcctcc tgttcaagac
     6061 gacgcgaacg ctccacggcg gccgatggcg cgggcagggc agggggagcc agttgcacgc
     6121 tgtcgcgctc gatcttggcc gtagcttgct ggaccatcga gccgacggac tggaaggttt
     6181 cgcggggcgc acgcatgacg gtgcggcttg cgatggtttc ggcatcctcg gcggaaaacc
     6241 ccgcgtcgat cagttcttgc ctgtatgcct tccggtcaaa cgtccgattc attcaccctc
     6301 cttgcgggat tgccccgact cacgccgggg caatgtgccc ttattcctga tttgacccgc
     6361 ctggtgcctt ggtgtccaga taatccacct tatcggcaat gaagtcggtc ccgtagaccg
     6421 tctggccgtc cttctcgtac ttggtattcc gaatcttgcc ctgcacgaat accagcgacc
     6481 ccttgcccaa atacttgccg tgggcctcgg cctgagagcc aaaacacttg atgcggaaga
     6541 agtcggtgcg ctcctgcttg tcgccggcat cgttgcgcca catctaggat ctgccaggaa
     6601 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     6661 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     6721 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     6781 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     6841 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     6901 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     6961 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     7021 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     7081 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     7141 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     7201 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     7261 cgaaaactca cgttaaggga ttttggtcat gaga
//
