LOCUS       sequence-349543-        6531 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 4, transcriptional unit for transient 
            expression of tryptophan decarboxylase (CrTDC) 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 6531)
  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 6531)
  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: pEPQD1CB0808
FEATURES             Location/Qualifiers
     source          1..6531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    131..164
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    190..223
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     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)"
     promoter        362..408
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    498..553
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     terminator      2077..2277
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    2323..2347
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2503..3651)
                     /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(3987..4575)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4067..4086)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4746..5537,5538..5606))
                     /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     5369..5388
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5607..5711)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5798..6429
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    join(6521..6531,1..14)
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
ORIGIN
        1 attgtggtgt aaacaaattg acgcttagac aacttaataa cacattgcgg acgtttttaa
       61 tgtactgggg tggatgcagt gggccccact ctgtgaagac aattacgaat tcccatgggg
      121 agcggtgcgg gtgccagggc gtgcccttgg gctccccggg cgcgtactcc actccccgag
      181 ccggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca ctccccccat
      241 ggtgcggtgc gggtgccagg gcgtgccctt gggctccccg ggcgcgtact ccactccccg
      301 agccggtgcg ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc cactcccctt
      361 cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggact ccggtatttt
      421 tacaacaatt accacaacaa aacaaacaac aaacaacatt acaatttact attctagtcg
      481 aatactgtat ttttacaaca attaccaaca acaacaaaca acaaacaaca ttacaattac
      541 tatttacaat tacaatgggc agcattgatt caacaaatgt agccatgtcc aattctccag
      601 ttggagaatt taagccactt gaagctgagg aattccgaaa acaagcccat cgtatggtag
      661 atttcatagc cgattattac aaaaatgtgg aaacatatcc ggtccttagc gaagtcgaac
      721 ctggatatct ccgaaaacgt atccccgaaa ccgctcctta cctccccgaa ccacttgacg
      781 acatcatgaa agatattcag aaggatatta tcccaggaat gacaaattgg atgagcccta
      841 atttttatgc attttttcct gccactgtta gttcagctgc ctttttagga gaaatgttgt
      901 ctactgccct aaattcagta ggctttactt gggtttcttc accagccgcc accgaattag
      961 aaatgattgt tatggattgg ttggctcaga tccttaaact ccccaaatct ttcatgtttt
     1021 caggtaccgg tggcggcgtc atccaaaaca ccactagcga gtccattctt tgtacaatca
     1081 ttgccgcccg ggaaagggcc ctggagaagc tcggtcccga tagtattgga aaacttgtct
     1141 gttacggatc cgatcaaacc cataccatgt tccccaaaac ttgcaaattg gcgggaattt
     1201 atccgaataa tattaggtta atacctacga ccgtcgaaac ggatttcggc atctcacctc
     1261 aagttctacg aaaaatggtc gaggatgacg tggcggccgg atatgtaccg ctgttcttat
     1321 gcgctaccct gggtaccacc tcgaccacgg ctaccgatcc tgtggactca ctttctgaaa
     1381 tcgctaacga gtttggtatt tggatccacg tggatgctgc ttatgcggga agcgcctgta
     1441 tatgtcccga gtttagacat tacttggatg gaatcgaacg agttgactca ctgagtctga
     1501 gtccacacaa atggctactc gcttacttag attgcacttg cttgtgggtc aagcaaccac
     1561 atttgttact aagggcactc actacgaatc ctgagtattt aaaaaataaa cagagtgatt
     1621 tagacaaagt tgtggacttc aaaaattggc aaatcgcaac gggacgaaaa tttcggtcgc
     1681 tgaaactttg gctcatttta cgtagctatg gagttgttaa tttacagagt catattcgtt
     1741 ctgacgtcgc aatgggcaaa atgttcgaag aatgggttag atcagactcc agattcgaaa
     1801 ttgtggtacc gagaaacttt tctcttgttt gttttagatt aaaacctgac gtttcgagtt
     1861 tacatgtaga agaagtgaat aagaaacttt tggacatgct taactcgacg ggacgagttt
     1921 atatgactca tactattgtg ggaggcatat acatgctaag actggctgtt ggctcgtcac
     1981 taactgaaga acatcatgta cgccgtgttt gggatttgat tcaaaaatta accgatgatt
     2041 tgctcaaaga agcttgagct tgatcccccg tcgacagcta gctatatcat caatttatgt
     2101 attacacata atatcgcact cagtctttca tctacggcaa tgtaccagct gatataatca
     2161 gttattgaaa tatttctgaa tttaaacttg catcaataaa tttatgtttt tgcttggact
     2221 ataatacctg acttgttatt ttatcaataa atatttaaac tatatttctt tcaagatcgc
     2281 tcagattgtc ttctgcacga agtggtttaa actatcagtg tttgacagga tatattggcg
     2341 ggtaaaccta agagaaaaga gcgtttatta gaataatcgg atatttaaaa gggcgtgaaa
     2401 aggtttatcc gttcgtccat ttgtatgtgc atgccaacca cagggttccc cagatcaggc
     2461 gctggctgct gaacccccag ccggaactga ccccacaagg ccctagcgtt tgcaatgcac
     2521 caggtcatca ttgacccagg cgtgttccac caggccgctg cctcgcaact cttcgcaggc
     2581 ttcgccgacc tgctcgcgcc acttcttcac gcgggtggaa tccgatccgc acatgaggcg
     2641 gaaggtttcc agcttgagcg ggtacggctc ccggtgcgag ctgaaatagt cgaacatccg
     2701 tcgggccgtc ggcgacagct tgcggtactt ctcccatatg aatttcgtgt agtggtcgcc
     2761 agcaaacagc acgacgattt cctcgtcgat caggacctgg caacgggacg ttttcttgcc
     2821 acggtccagg acgcggaagc ggtgcagcag cgacaccgat tccaggtgcc caacgcggtc
     2881 ggacgtgaag cccatcgccg tcgcctgtag gcgcgacagg cattcctcgg ccttcgtgta
     2941 ataccggcca ttgatcgacc agcccaggtc ctggcaaagc tcgtagaacg tgaaggtgat
     3001 cggctcgccg ataggggtgc gcttcgcgta ctccaacacc tgctgccaca ccagttcgtc
     3061 atcgtcggcc cgcagctcga cgccggtgta ggtgatcttc acgtccttgt tgacgtggaa
     3121 aatgaccttg ttttgcagcg cctcgcgcgg gattttcttg ttgcgcgtgg tgaacagggc
     3181 agagcgggcc gtgtcgtttg gcatcgctcg catcgtgtcc ggccacggcg caatatcgaa
     3241 caaggaaagc tgcatttcct tgatctgctg cttcgtgtgt ttcagcaacg cggcctgctt
     3301 ggcctcgctg acctgttttg ccaggtcctc gccggcggtt tttcgcttct tggtcgtcat
     3361 agttcctcgc gtgtcgatgg tcatcgactt cgccaaacct gccgcctcct gttcaagacg
     3421 acgcgaacgc tccacggcgg ccgatggcgc gggcagggca gggggagcca gttgcacgct
     3481 gtcgcgctcg atcttggccg tagcttgctg gaccatcgag ccgacggact ggaaggtttc
     3541 gcggggcgca cgcatgacgg tgcggcttgc gatggtttcg gcatcctcgg cggaaaaccc
     3601 cgcgtcgatc agttcttgcc tgtatgcctt ccggtcaaac gtccgattca ttcaccctcc
     3661 ttgcgggatt gccccgactc acgccggggc aatgtgccct tattcctgat ttgacccgcc
     3721 tggtgccttg gtgtccagat aatccacctt atcggcaatg aagtcggtcc cgtagaccgt
     3781 ctggccgtcc ttctcgtact tggtattccg aatcttgccc tgcacgaata ccagcgaccc
     3841 cttgcccaaa tacttgccgt gggcctcggc ctgagagcca aaacacttga tgcggaagaa
     3901 gtcggtgcgc tcctgcttgt cgccggcatc gttgcgccac atctaggatc tgccaggaac
     3961 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4021 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4081 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4141 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4201 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4261 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4321 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4381 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     4441 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4501 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4561 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4621 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     4681 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     4741 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     4801 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     4861 ggccccagtg ctgcaatgat accgcgagaa ccacgctcac cggctccaga tttatcagca
     4921 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     4981 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5041 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5101 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5161 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5221 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5281 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5341 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5401 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5461 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     5521 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     5581 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     5641 caaggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     5701 ggggttccgc gcacgaattg gccagcgctg ccatttttgg ggtgaggccg ttcgcggccg
     5761 aggggcgcag cccctggggg gatgggaggc ccgcgttagc gggccgggag ggttcgagaa
     5821 gggggggcac cccccttcgg cgtgcgcggt cacgcgcaca gggcgcagcc ctggttaaaa
     5881 acaaggttta taaatattgg tttaaaagca ggttaaaaga caggttagcg gtggccgaaa
     5941 aacgggcgga aacccttgca aatgctggat tttctgcctg tggacagccc ctcaaatgtc
     6001 aataggtgcg cccctcatct gtcagcactc tgcccctcaa gtgtcaagga tcgcgcccct
     6061 catctgtcag tagtcgcgcc cctcaagtgt caataccgca gggcacttat ccccaggctt
     6121 gtccacatca tctgtgggaa actcgcgtaa aatcaggcgt tttcgccgat ttgcgaggct
     6181 ggccagctcc acgtcgccgg ccgaaatcga gcctgcccct catctgtcaa cgccgcgccg
     6241 ggtgagtcgg cccctcaagt gtcaacgtcc gcccctcatc tgtcagtgag ggccaagttt
     6301 tccgcgaggt atccacaacg ccggcggccg cggtgtctcg cacacggctt cgacggcgtt
     6361 tctggcgcgt ttgcagggcc atagacggcc gccagcccag cggcgagggc aaccagcccg
     6421 gtgagcgtcg caaaggagat cctgatctga ctgatgggct gcctgtatcg agtggtgatt
     6481 ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata t
//
