LOCUS       sequence-349544-        6075 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 1, transcriptional unit for transient 
            expression of strictosidine synthase (CrSTR) 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 6075)
  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 6075)
  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: pEPQD1CB0802
FEATURES             Location/Qualifiers
     source          1..6075
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      99..730
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    822..846
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     protein_bind    963..996
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    1022..1055
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    1085..1118
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     protein_bind    1144..1177
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     promoter        1194..1240
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    1330..1385
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5'-leader sequence (Gallie et al., 1988)"
     polyA_signal    2480..2654
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    2699..2723
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(2879..4027)
                     /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(4363..4951)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4443..4462)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5122..5913,5914..5982))
                     /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     5745..5764
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(5983..6075,1..12))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aggggttccg cgcacgaatt ggccagcgct gccatttttg gggtgaggcc gttcgcggcc
       61 gaggggcgca gcccctgggg ggatgggagg cccgcgttag cgggccggga gggttcgaga
      121 agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac agggcgcagc cctggttaaa
      181 aacaaggttt ataaatattg gtttaaaagc aggttaaaag acaggttagc ggtggccgaa
      241 aaacgggcgg aaacccttgc aaatgctgga ttttctgcct gtggacagcc cctcaaatgt
      301 caataggtgc gcccctcatc tgtcagcact ctgcccctca agtgtcaagg atcgcgcccc
      361 tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc agggcactta tccccaggct
      421 tgtccacatc atctgtggga aactcgcgta aaatcaggcg ttttcgccga tttgcgaggc
      481 tggccagctc cacgtcgccg gccgaaatcg agcctgcccc tcatctgtca acgccgcgcc
      541 gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga gggccaagtt
      601 ttccgcgagg tatccacaac gccggcggcc gcggtgtctc gcacacggct tcgacggcgt
      661 ttctggcgcg tttgcagggc catagacggc cgccagccca gcggcgaggg caaccagccc
      721 ggtgagcgtc gcaaaggaga tcctgatctg actgatgggc tgcctgtatc gagtggtgat
      781 tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat atattgtggt
      841 gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt aatgtactgg
      901 ggtggatgca gtgggcccca ctctgtgaag acaatgccga attcggatcc ggagcggtgc
      961 gggtgccagg gcgtgccctt gggctccccg ggcgcgtact ccactccccg agccggtgcg
     1021 ggtgccaggg cgtgcccttg ggctccccgg gcgcgtactc cactcccccc atggtgcggt
     1081 gcgggtgcca gggcgtgccc ttgggctccc cgggcgcgta ctccactccc cgagccggtg
     1141 cgggtgccag ggcgtgccct tgggctcccc gggcgcgtac tccactcccc ttcgcaagac
     1201 ccttcctcta tataaggaag ttcatttcat ttggagagga ctccggtatt tttacaacaa
     1261 ttaccacaac aaaacaaaca acaaacaaca ttacaattta ctattctagt cgaatactgt
     1321 atttttacaa caattaccaa caacaacaaa caacaaacaa cattacaatt actatttaca
     1381 attacaatgg caaacttttc tgaatctaaa tccatgatgg cagttttctt catgtttttc
     1441 ctgcttcttt tgtcatcttc tagttcttca tcttcaagtt caccaatttt gaaaaagatt
     1501 tttattgaaa gcccttccta tgctccgaat gccttcacct tcgattcaac tgataaaggg
     1561 ttctacactt ccgtccaaga tggccgagtt atcaaatatg aagggccaaa ttcaggcttc
     1621 actgacttcg cctacgcatc tcccttctgg aacaaagctt tttgtgagaa cagcaccgat
     1681 ccagagaaaa gaccattgtg tgggaggaca tatgatattt cctatgacta taagaacagc
     1741 caaatgtaca ttgttgatgg ccattaccat ctttgtgtgg ttggaaaaga aggtgggtat
     1801 gccacacaac tagccacaag tgtgcaagga gtgccattca aatggctcta tgcagtaact
     1861 gttgatcaga gaacagggat tgtttatttc actgatgtta gctccataca tgatgacagt
     1921 cccgaaggtg tggaagaaat catgaataca agtgatagaa cagggagatt aatgaagtat
     1981 gatccttcaa caaaagaaac caccttatta ttgaaagagc tacatgttcc cggcggtgca
     2041 gaaatcagcg cagatggttc ctttgttgta gtagcagaat ttttaagcaa tcggatagtg
     2101 aagtattggc tagaagggcc aaagaaaggc agtgcagagt tcttagttac aatcccaaat
     2161 ccaggaaata taaagaggaa ttctgatggc catttttggg tgtcatctag tgaagaatta
     2221 gatggaggtc aacatggaag agttgtttca agaggaatta agtttgatgg atttgggaat
     2281 attcttcaag ttataccact tccaccacca tatgaaggtg aacattttga acagattcaa
     2341 gagcacgatg gtttgttata cattggaagt ctcttccata gctctgtggg tatattagtg
     2401 tatgatgatc atgataacaa gggaaattct tatgtttcta gctgagcttc tctagctaga
     2461 gtcgatcgac aagctcgagt ttctccataa taatgtgtga gtagttccca gataagggaa
     2521 ttagggttcc tatagggttt cgctcatgtg ttgagcatat aagaaaccct tagtatgtat
     2581 ttgtatttgt aaaatacttc tatcaataaa atttctaatt cctaaaacca aaatccagta
     2641 ctaaaatcca gatcgctgca attgtcttct gcacgaagtg gtttaaacta tcagtgtttg
     2701 acaggatata ttggcgggta aacctaagag aaaagagcgt ttattagaat aatcggatat
     2761 ttaaaagggc gtgaaaaggt ttatccgttc gtccatttgt atgtgcatgc caaccacagg
     2821 gttccccaga tcaggcgctg gctgctgaac ccccagccgg aactgacccc acaaggccct
     2881 agcgtttgca atgcaccagg tcatcattga cccaggcgtg ttccaccagg ccgctgcctc
     2941 gcaactcttc gcaggcttcg ccgacctgct cgcgccactt cttcacgcgg gtggaatccg
     3001 atccgcacat gaggcggaag gtttccagct tgagcgggta cggctcccgg tgcgagctga
     3061 aatagtcgaa catccgtcgg gccgtcggcg acagcttgcg gtacttctcc catatgaatt
     3121 tcgtgtagtg gtcgccagca aacagcacga cgatttcctc gtcgatcagg acctggcaac
     3181 gggacgtttt cttgccacgg tccaggacgc ggaagcggtg cagcagcgac accgattcca
     3241 ggtgcccaac gcggtcggac gtgaagccca tcgccgtcgc ctgtaggcgc gacaggcatt
     3301 cctcggcctt cgtgtaatac cggccattga tcgaccagcc caggtcctgg caaagctcgt
     3361 agaacgtgaa ggtgatcggc tcgccgatag gggtgcgctt cgcgtactcc aacacctgct
     3421 gccacaccag ttcgtcatcg tcggcccgca gctcgacgcc ggtgtaggtg atcttcacgt
     3481 ccttgttgac gtggaaaatg accttgtttt gcagcgcctc gcgcgggatt ttcttgttgc
     3541 gcgtggtgaa cagggcagag cgggccgtgt cgtttggcat cgctcgcatc gtgtccggcc
     3601 acggcgcaat atcgaacaag gaaagctgca tttccttgat ctgctgcttc gtgtgtttca
     3661 gcaacgcggc ctgcttggcc tcgctgacct gttttgccag gtcctcgccg gcggtttttc
     3721 gcttcttggt cgtcatagtt cctcgcgtgt cgatggtcat cgacttcgcc aaacctgccg
     3781 cctcctgttc aagacgacgc gaacgctcca cggcggccga tggcgcgggc agggcagggg
     3841 gagccagttg cacgctgtcg cgctcgatct tggccgtagc ttgctggacc atcgagccga
     3901 cggactggaa ggtttcgcgg ggcgcacgca tgacggtgcg gcttgcgatg gtttcggcat
     3961 cctcggcgga aaaccccgcg tcgatcagtt cttgcctgta tgccttccgg tcaaacgtcc
     4021 gattcattca ccctccttgc gggattgccc cgactcacgc cggggcaatg tgcccttatt
     4081 cctgatttga cccgcctggt gccttggtgt ccagataatc caccttatcg gcaatgaagt
     4141 cggtcccgta gaccgtctgg ccgtccttct cgtacttggt attccgaatc ttgccctgca
     4201 cgaataccag cgaccccttg cccaaatact tgccgtgggc ctcggcctga gagccaaaac
     4261 acttgatgcg gaagaagtcg gtgcgctcct gcttgtcgcc ggcatcgttg cgccacatct
     4321 aggatctgcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4381 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4441 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4501 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4561 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4621 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4681 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4741 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4801 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4861 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4921 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4981 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5041 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5101 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5161 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     5221 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagaaccac gctcaccggc
     5281 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     5341 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     5401 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     5461 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     5521 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     5581 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     5641 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     5701 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     5761 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     5821 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     5881 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     5941 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6001 ttattgaagc atttatcaag gttattgtct catgagcgga tacatatttg aatgtattta
     6061 gaaaaataaa caaat
//
