LOCUS       sequence-349450-        6834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 4, transcriptional unit for transient 
            expression of geraniol synthase (CrGES) 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 6834)
  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 6834)
  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: pEPQD1CB0787
FEATURES             Location/Qualifiers
     source          1..6834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(204..995,996..1064))
                     /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     827..846
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1065..1169)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1256..1887
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    1979..2003
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    2120..2153
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    2179..2212
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    2242..2275
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    2301..2334
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        2351..2397
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     CDS             4048..4059
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     terminator      4369..4569
                     /label=gene 7 terminator
                     /note="transcription terminator for octopine-type Ti 
                     plasmid gene 7"
     misc_feature    4615..4639
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(4795..5943)
                     /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(join(6279..6834,1..33))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6359..6378)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
       61 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
      121 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
      181 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
      241 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
      301 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaacc acgctcaccg
      361 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
      421 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
      481 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
      541 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
      601 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
      661 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
      721 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
      781 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
      841 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
      901 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
      961 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     1021 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     1081 tattattgaa gcatttatca aggttattgt ctcatgagcg gatacatatt tgaatgtatt
     1141 tagaaaaata aacaaatagg ggttccgcgc acgaattggc cagcgctgcc atttttgggg
     1201 tgaggccgtt cgcggccgag gggcgcagcc cctgggggga tgggaggccc gcgttagcgg
     1261 gccgggaggg ttcgagaagg gggggcaccc cccttcggcg tgcgcggtca cgcgcacagg
     1321 gcgcagccct ggttaaaaac aaggtttata aatattggtt taaaagcagg ttaaaagaca
     1381 ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa tgctggattt tctgcctgtg
     1441 gacagcccct caaatgtcaa taggtgcgcc cctcatctgt cagcactctg cccctcaagt
     1501 gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc tcaagtgtca ataccgcagg
     1561 gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac tcgcgtaaaa tcaggcgttt
     1621 tcgccgattt gcgaggctgg ccagctccac gtcgccggcc gaaatcgagc ctgcccctca
     1681 tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt caacgtccgc ccctcatctg
     1741 tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc ggcggccgcg gtgtctcgca
     1801 cacggcttcg acggcgtttc tggcgcgttt gcagggccat agacggccgc cagcccagcg
     1861 gcgagggcaa ccagcccggt gagcgtcgca aaggagatcc tgatctgact gatgggctgc
     1921 ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg ctggctggtg
     1981 gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac acattgcgga
     2041 cgtttttaat gtactggggt ggatgcagtg ggccccactc tgtgaagaca attacgaatt
     2101 cccatgggga gcggtgcggg tgccagggcg tgcccttggg ctccccgggc gcgtactcca
     2161 ctccccgagc cggtgcgggt gccagggcgt gcccttgggc tccccgggcg cgtactccac
     2221 tccccccatg gtgcggtgcg ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc
     2281 cactccccga gccggtgcgg gtgccagggc gtgcccttgg gctccccggg cgcgtactcc
     2341 actccccttc gcaagaccct tcctctatat aaggaagttc atttcatttg gagaggactc
     2401 cggtattttt acaacaatta ccacaacaaa acaaacaaca aacaacatta caatttacta
     2461 ttctagtcga atactgtata agagctcatt tttacaacaa ttaccaacaa caacaaacaa
     2521 caaacaacat tacaattaca tttacaatta tcgataacat ggcttcttct atgctttctt
     2581 ctgctgctgt tgttgctact cgtgctagtg ctgctcaagc tagtatggtt gctcctttta
     2641 ctggacttaa gtctgctgct tcttttcctg ttactagaaa gcaaaacaac cttgatatta
     2701 cttctattgc tagtaacgga ggaagagttc gagcaatgtc tctgcctttg gcaactccat
     2761 tgatcaaaga caatgaatct ctcatcaagt tcttgcgcca acccctggtg cttcctcatg
     2821 aggttgatga cagcacaaaa aggagggaat tgttggaaag aacaagaaaa gaactagaat
     2881 taaatgcaga aaaaccattg gaggccttga agatgataga tataattcaa agattgggat
     2941 tatcatatca ttttgaagat gatattaatt caattctcac aggattttca aatattagca
     3001 gccaaactca tgaagatctc ctcactgctt cactttgttt tcgattgctt cgacacaatg
     3061 ggcataagat caatcctgat atattccaaa aattcatgga caacaatgga aagtttaaag
     3121 attcattaaa ggatgacaca ttaggcatgt taagcttata tgaagcttca tatttgggag
     3181 ccaatggaga agaaatattg atggaagccc aagaattcac caaaactcac ctgaaaaact
     3241 cattgccagc catggcacca tctctttcta agaaggtttc tcaagcttta gagcaaccaa
     3301 gacatagaag aatgttgagg ttagaagcta gaagatttat tgaagaatat ggtgctgaaa
     3361 atgaccataa tccagacctt cttgagcttg caaaattgga ttataacaaa gtccaatctc
     3421 tacaccaaat ggaattgtct gagataacaa ggtggtggaa acaattaggg cttgtggata
     3481 aactcacctt tgctcgagat cgaccccttg aatgctttct ttggacagtg ggattattac
     3541 cagagcctaa gtattcaggt tgcagaattg agcttgcaaa aaccatagcc attttgcttg
     3601 tcattgatga tatctttgat actcatggta ccctagatga gcttcttcta ttcactaatg
     3661 ccattaaaag atgggatctt gaggccatgg aagatttacc agaatatatg agaatttgtt
     3721 acatggcatt gtacaatact actaatgaaa tttgctataa agttcttaag gaaaatggtt
     3781 ggagtgttct tccttaccta aaggcaacgt ggattgatat gattgaagga ttcatggttg
     3841 aagcagaatg gttcaattct gattatgtac caaacatgga agaatatgta gaaaatggag
     3901 ttagaacagc aggatcatat atggccttag tccatttgtt ctttctaata gggcaaggtg
     3961 tcactgaaga taatgtgaaa ttactgatta aaccctatcc aaagctcttt tcctcctcag
     4021 gaagaatcct tcgcctttgg gatgatttgg gaactgcaaa ggaggaacaa gaaagaggag
     4081 acttggcatc aagcatacaa ttgtttatga gagagaaaga gataaaatca gaagaagaag
     4141 gaaggaaagg aatattggaa attatagaga atttatggaa agaattgaat ggagaattag
     4201 tttatagaga agaaatgcct cttgcaataa tcaagacagc attcaacatg gcaagagctt
     4261 cccaagttgt gtatcaacat gaagaggaca cctatttttc aagtgtagat aattatgtaa
     4321 aagctttgtt ttttacacct tgtttttaag cttgatcccc cgtcgacagc tagctatatc
     4381 atcaatttat gtattacaca taatatcgca ctcagtcttt catctacggc aatgtaccag
     4441 ctgatataat cagttattga aatatttctg aatttaaact tgcatcaata aatttatgtt
     4501 tttgcttgga ctataatacc tgacttgtta ttttatcaat aaatatttaa actatatttc
     4561 tttcaagatc gctcagattg tcttctgcac gaagtggttt aaactatcag tgtttgacag
     4621 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataatc ggatatttaa
     4681 aagggcgtga aaaggtttat ccgttcgtcc atttgtatgt gcatgccaac cacagggttc
     4741 cccagatcag gcgctggctg ctgaaccccc agccggaact gaccccacaa ggccctagcg
     4801 tttgcaatgc accaggtcat cattgaccca ggcgtgttcc accaggccgc tgcctcgcaa
     4861 ctcttcgcag gcttcgccga cctgctcgcg ccacttcttc acgcgggtgg aatccgatcc
     4921 gcacatgagg cggaaggttt ccagcttgag cgggtacggc tcccggtgcg agctgaaata
     4981 gtcgaacatc cgtcgggccg tcggcgacag cttgcggtac ttctcccata tgaatttcgt
     5041 gtagtggtcg ccagcaaaca gcacgacgat ttcctcgtcg atcaggacct ggcaacggga
     5101 cgttttcttg ccacggtcca ggacgcggaa gcggtgcagc agcgacaccg attccaggtg
     5161 cccaacgcgg tcggacgtga agcccatcgc cgtcgcctgt aggcgcgaca ggcattcctc
     5221 ggccttcgtg taataccggc cattgatcga ccagcccagg tcctggcaaa gctcgtagaa
     5281 cgtgaaggtg atcggctcgc cgataggggt gcgcttcgcg tactccaaca cctgctgcca
     5341 caccagttcg tcatcgtcgg cccgcagctc gacgccggtg taggtgatct tcacgtcctt
     5401 gttgacgtgg aaaatgacct tgttttgcag cgcctcgcgc gggattttct tgttgcgcgt
     5461 ggtgaacagg gcagagcggg ccgtgtcgtt tggcatcgct cgcatcgtgt ccggccacgg
     5521 cgcaatatcg aacaaggaaa gctgcatttc cttgatctgc tgcttcgtgt gtttcagcaa
     5581 cgcggcctgc ttggcctcgc tgacctgttt tgccaggtcc tcgccggcgg tttttcgctt
     5641 cttggtcgtc atagttcctc gcgtgtcgat ggtcatcgac ttcgccaaac ctgccgcctc
     5701 ctgttcaaga cgacgcgaac gctccacggc ggccgatggc gcgggcaggg cagggggagc
     5761 cagttgcacg ctgtcgcgct cgatcttggc cgtagcttgc tggaccatcg agccgacgga
     5821 ctggaaggtt tcgcggggcg cacgcatgac ggtgcggctt gcgatggttt cggcatcctc
     5881 ggcggaaaac cccgcgtcga tcagttcttg cctgtatgcc ttccggtcaa acgtccgatt
     5941 cattcaccct ccttgcggga ttgccccgac tcacgccggg gcaatgtgcc cttattcctg
     6001 atttgacccg cctggtgcct tggtgtccag ataatccacc ttatcggcaa tgaagtcggt
     6061 cccgtagacc gtctggccgt ccttctcgta cttggtattc cgaatcttgc cctgcacgaa
     6121 taccagcgac cccttgccca aatacttgcc gtgggcctcg gcctgagagc caaaacactt
     6181 gatgcggaag aagtcggtgc gctcctgctt gtcgccggca tcgttgcgcc acatctagga
     6241 tctgccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     6301 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     6361 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     6421 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     6481 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     6541 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     6601 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     6661 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     6721 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     6781 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caag
//
