LOCUS       sequence-349530-        8131 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Level 1, position 1, transcriptional unit for transient 
            expression of gal4AD-phiC31 driven by 35S promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8131)
  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 8131)
  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: pEPQD1CB0905
FEATURES             Location/Qualifiers
     source          1..8131
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(404..1195,1196..1264))
                     /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     1027..1046
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1265..1369)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1456..2087
                     /label=oriV
                     /note="incP origin of replication"
     misc_feature    2179..2203
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        3301..3646
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     3589..3609
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     CDS             4020..5834
                     /codon_start=1
                     /product="integrase from phage phi-C31"
                     /label=phage phi-C31 integrase
                     /translation="MDTYAGAYDRQSRERENSSAASPATQRSANEDKAADLQREVERDG
                     GRFRFVGHFSEAPGTSAFGTAERPEFERILNECRAGRLNMIIVYDVSRFSRLKVMDAIP
                     IVSELLALGVTIVSTQEGVFRQGNVMDLIHLIMRLDASHKESSLKSAKILDTKNLQREL
                     GGYVGGKAPYGFELVSETKEITRNGRMVNVVINKLAHSTTPLTGPFEFEPDVIRWWWRE
                     IKTHKHLPFKPGSQAAIHPGSITGLCKRMDADAVPTRGETIGKKTASSAWDPATVMRIL
                     RDPRIAGFAAEVIYKKKPDGTPTTKIEGYRIQRDPITLRPVELDCGPIIEPAEWYELQA
                     WLDGRGRGKGLSRGQAILSAMDKLYCECGAVMTSKRGEESIKDSYRCRRRKVVDPSAPG
                     QHEGTCNVSMAALDKFVAERIFNKIRHAEGDEETLALLWEAARRFGKLTEAPEKSGERA
                     NLVAERADALNALEELYEDRAAGAYDGPVGRKHFRKQQAALTLRQQGAEERLAELEAAE
                     APKLPLDQWFPEDADADPTGPKSWWGRASVDDKRVFVGLFVDKIVVTKSTTGRGQGTPI
                     EKRASITWAKPPTDDDEDDAQDGTEDVAA"
     CDS             5835..5855
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    5893..6067
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     misc_feature    6112..6136
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(6292..7440)
                     /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(7776..8131,1..233))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7856..7875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
       61 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      121 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      181 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      241 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
      301 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
      361 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
      421 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
      481 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
      541 cgcgagaacc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
      601 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
      661 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
      721 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
      781 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
      841 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
      901 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
      961 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     1021 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     1081 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     1141 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     1201 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     1261 tcatactctt cctttttcaa tattattgaa gcatttatca aggttattgt ctcatgagcg
     1321 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acgaattggc
     1381 cagcgctgcc atttttgggg tgaggccgtt cgcggccgag gggcgcagcc cctgggggga
     1441 tgggaggccc gcgttagcgg gccgggaggg ttcgagaagg gggggcaccc cccttcggcg
     1501 tgcgcggtca cgcgcacagg gcgcagccct ggttaaaaac aaggtttata aatattggtt
     1561 taaaagcagg ttaaaagaca ggttagcggt ggccgaaaaa cgggcggaaa cccttgcaaa
     1621 tgctggattt tctgcctgtg gacagcccct caaatgtcaa taggtgcgcc cctcatctgt
     1681 cagcactctg cccctcaagt gtcaaggatc gcgcccctca tctgtcagta gtcgcgcccc
     1741 tcaagtgtca ataccgcagg gcacttatcc ccaggcttgt ccacatcatc tgtgggaaac
     1801 tcgcgtaaaa tcaggcgttt tcgccgattt gcgaggctgg ccagctccac gtcgccggcc
     1861 gaaatcgagc ctgcccctca tctgtcaacg ccgcgccggg tgagtcggcc cctcaagtgt
     1921 caacgtccgc ccctcatctg tcagtgaggg ccaagttttc cgcgaggtat ccacaacgcc
     1981 ggcggccgcg gtgtctcgca cacggcttcg acggcgtttc tggcgcgttt gcagggccat
     2041 agacggccgc cagcccagcg gcgagggcaa ccagcccggt gagcgtcgca aaggagatcc
     2101 tgatctgact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg
     2161 gagctgttgg ctggctggtg gcaggatata ttgtggtgta aacaaattga cgcttagaca
     2221 acttaataac acattgcgga cgtttttaat gtactggggt ggatgcagtg ggccccactc
     2281 tgtgaagaca atgccgaatt cggatccgga ggaattccaa tcccacaaaa atctgagctt
     2341 aacagcacag ttgctcctct cagagcagaa tcgggtattc aacaccctca tatcaactac
     2401 tacgttgtgt ataacggtcc acatgccggt atatacgatg actggggttg tacaaaggcg
     2461 gcaacaaacg gcgttcccgg agttgcacac aagaaatttg ccactattac agaggcaaga
     2521 gcagcagctg acgcgtacac aacaagtcag caaacagaca ggttgaactt catccccaaa
     2581 ggagaagctc aactcaagcc caagagcttt gctaaggccc taacaagccc accaaagcaa
     2641 aaagcccact ggctcacgct aggaaccaaa aggcccagca gtgatccagc cccaaaagag
     2701 atctcctttg ccccggagat tacaatggac gatttcctct atctttacga tctaggaagg
     2761 aagttcgaag gtgaaggtga cgacactatg ttcaccactg ataatgagaa ggttagcctc
     2821 ttcaatttca gaaagaatgc tgacccacag atggttagag aggcctacgc agcaagtctc
     2881 atcaagacga tctacccgag taacaatctc caggagatca aataccttcc caagaaggtt
     2941 aaagatgcag tcaaaagatt caggactaat tgcatcaaga acacagagaa agacatattt
     3001 ctcaagatca gaagtactat tccagtatgg acgattcaag gcttgcttca taaaccaagg
     3061 caagtaatag agattggagt ctctaaaaag gtagttccta ctgaatctaa ggccatgcat
     3121 ggagtctaag attcaaatcg aggatctaac agaactcgcc gtcaagactg gcgaacagtt
     3181 catacagagt cttttacgac tcaatgacaa gaagaaaatc ttcgtcaaca tggtggagca
     3241 cgacactctg gtctactcca aaaatgtcaa agatacagtc tcagaagatc aaagggctat
     3301 tgagactttt caacaaagga taatttcggg aaacctcctc ggattccatt gcccagctat
     3361 ctgtcacttc atcgaaagga cagtagaaaa ggaaggtggc tcctacaaat gccatcattg
     3421 cgataaagga aaggctatca ttcaagatct ctctgccgac agtggtccca aagatggacc
     3481 cccacccacg aggagcatcg tggaaaaaga agaggttcca accacgtcta caaagcaagt
     3541 ggattgatgt gacatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca
     3601 agacccttcc tctatataag gaagttcatt tcatttggag aggacacgct cgagtataag
     3661 agctcatttt tacaacaatt accaacaaca acaaacaaca aacaacatta caattacatt
     3721 tacaattatc gatacccatg acaaattctc aagcgctttc acaaccaatt gcctcctcta
     3781 acgttcatga taacttcatg aataatgaaa tcacggctag taaaattgat gatggtaata
     3841 attcaaaacc actgtcacct ggttggacgg accaaactgc gtataacgcg tttggaatca
     3901 ctacagggat gtttaatacc actacaatgg atgatgtata taactatcta ttcgatgatg
     3961 aagatacccc accaaaacca aaaaaagagg gtggcagtgg agggggcgga agcgggggaa
     4021 tggacacgta tgccggcgct tatgatagac aatcaagaga aagagagaac tcttctgccg
     4081 cctctccagc tacacagaga tctgccaacg aggataaagc tgctgattta caaagagagg
     4141 tagagagaga tgggggtagg tttagatttg tgggtcattt ctctgaggct cctggtacat
     4201 cagcctttgg tactgctgaa cggcctgaat ttgaaagaat attaaatgag tgtcgggccg
     4261 gtcgactaaa tatgataata gtttatgatg tatctaggtt cagtagactc aaagttatgg
     4321 atgcaattcc gatcgtgagc gagctacttg cactcggtgt tactatcgta agtacacaag
     4381 aaggtgtatt caggcagggt aatgtgatgg accttataca ccttattatg cgactcgacg
     4441 ctagccataa agagtcatca ctgaagagtg ctaaaatctt ggacaccaag aacctgcaaa
     4501 gggagcttgg aggctatgta ggaggaaaag ctccgtatgg tttcgaactc gtgagtgaga
     4561 caaaagagat taccagaaat ggtaggatgg tgaacgtggt tattaataag ctagctcata
     4621 gcacaacccc tctcacaggg ccttttgagt tcgaacctga cgttattaga tggtggtgga
     4681 gagagatcaa aactcacaaa catcttccat ttaagccagg tagccaggct gctatccacc
     4741 ctggatctat cactgggctt tgtaaaagga tggatgctga cgcagtgcca actcgagggg
     4801 aaacaatcgg taagaaaact gcatcttccg cttgggatcc cgctacagtt atgagaatcc
     4861 ttcgggatcc gcgtattgct ggattcgccg cagaagtcat ttataagaaa aaacctgatg
     4921 gtactcctac tacaaaaatc gaaggttata ggatccaaag ggatcctatt actttgcgtc
     4981 cagttgaatt agactgtgga ccgatcatcg aacctgccga gtggtacgag ttgcaggcct
     5041 ggcttgatgg tagagggcgt ggaaagggtt tatctcgggg acaggcaatt ttgtcagcaa
     5101 tggataagct ttattgtgag tgtggggctg ttatgacatc aaaacgtgga gaagagtcaa
     5161 tcaaggattc ttacaggtgc agacgtcgta aagtggtgga cccttctgct ccaggacagc
     5221 atgagggtac atgcaacgtt tctatggctg ctcttgataa atttgtggcc gaacgtattt
     5281 tcaacaagat tagacacgct gaaggggatg aggaaacttt agctcttctt tgggaggctg
     5341 cccggcgttt tggtaagcta acagaggcac cagaaaaaag tggcgaaaga gctaatttgg
     5401 tagccgaaag agcagacgcc ttgaatgcat tggaagaact ttacgaagat agggctgccg
     5461 gagcatatga tggaccagtt ggtcggaagc atttcaggaa gcaacaagct gcactaactt
     5521 tgagacagca aggcgcagag gagcgactcg cagagcttga ggccgccgag gccccaaagc
     5581 ttccactcga tcaatggttt cctgaggatg ccgacgctga tcctacaggt ccaaagagct
     5641 ggtggggaag agctagcgta gacgataaga gagtgtttgt gggtctattt gttgacaaga
     5701 ttgtagttac taagtccact acgggtcgag gacagggcac tcccatcgaa aagagagcca
     5761 gtattacctg ggcaaagcct ccaaccgatg atgacgagga cgatgcacaa gatggtaccg
     5821 aggatgtggc agcaccaaaa aaaaagagga aggtgtaagc ttctctagct agagtcgatc
     5881 gacaagctcg agtttctcca taataatgtg tgagtagttc ccagataagg gaattagggt
     5941 tcctataggg tttcgctcat gtgttgagca tataagaaac ccttagtatg tatttgtatt
     6001 tgtaaaatac ttctatcaat aaaatttcta attcctaaaa ccaaaatcca gtactaaaat
     6061 ccagatcgct gcaattgtct tctgcacgaa gtggtttaaa ctatcagtgt ttgacaggat
     6121 atattggcgg gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag
     6181 ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca tgccaaccac agggttcccc
     6241 agatcaggcg ctggctgctg aacccccagc cggaactgac cccacaaggc cctagcgttt
     6301 gcaatgcacc aggtcatcat tgacccaggc gtgttccacc aggccgctgc ctcgcaactc
     6361 ttcgcaggct tcgccgacct gctcgcgcca cttcttcacg cgggtggaat ccgatccgca
     6421 catgaggcgg aaggtttcca gcttgagcgg gtacggctcc cggtgcgagc tgaaatagtc
     6481 gaacatccgt cgggccgtcg gcgacagctt gcggtacttc tcccatatga atttcgtgta
     6541 gtggtcgcca gcaaacagca cgacgatttc ctcgtcgatc aggacctggc aacgggacgt
     6601 tttcttgcca cggtccagga cgcggaagcg gtgcagcagc gacaccgatt ccaggtgccc
     6661 aacgcggtcg gacgtgaagc ccatcgccgt cgcctgtagg cgcgacaggc attcctcggc
     6721 cttcgtgtaa taccggccat tgatcgacca gcccaggtcc tggcaaagct cgtagaacgt
     6781 gaaggtgatc ggctcgccga taggggtgcg cttcgcgtac tccaacacct gctgccacac
     6841 cagttcgtca tcgtcggccc gcagctcgac gccggtgtag gtgatcttca cgtccttgtt
     6901 gacgtggaaa atgaccttgt tttgcagcgc ctcgcgcggg attttcttgt tgcgcgtggt
     6961 gaacagggca gagcgggccg tgtcgtttgg catcgctcgc atcgtgtccg gccacggcgc
     7021 aatatcgaac aaggaaagct gcatttcctt gatctgctgc ttcgtgtgtt tcagcaacgc
     7081 ggcctgcttg gcctcgctga cctgttttgc caggtcctcg ccggcggttt ttcgcttctt
     7141 ggtcgtcata gttcctcgcg tgtcgatggt catcgacttc gccaaacctg ccgcctcctg
     7201 ttcaagacga cgcgaacgct ccacggcggc cgatggcgcg ggcagggcag ggggagccag
     7261 ttgcacgctg tcgcgctcga tcttggccgt agcttgctgg accatcgagc cgacggactg
     7321 gaaggtttcg cggggcgcac gcatgacggt gcggcttgcg atggtttcgg catcctcggc
     7381 ggaaaacccc gcgtcgatca gttcttgcct gtatgccttc cggtcaaacg tccgattcat
     7441 tcaccctcct tgcgggattg ccccgactca cgccggggca atgtgccctt attcctgatt
     7501 tgacccgcct ggtgccttgg tgtccagata atccacctta tcggcaatga agtcggtccc
     7561 gtagaccgtc tggccgtcct tctcgtactt ggtattccga atcttgccct gcacgaatac
     7621 cagcgacccc ttgcccaaat acttgccgtg ggcctcggcc tgagagccaa aacacttgat
     7681 gcggaagaag tcggtgcgct cctgcttgtc gccggcatcg ttgcgccaca tctaggatct
     7741 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     7801 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     7861 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     7921 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     7981 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     8041 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     8101 agacacgact tatcgccact ggcagcagcc a
//
