LOCUS       sequence-349002-        3945 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 CDS (with stop codon) part for MoClo assembly, AATG-GCTT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3945)
  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 3945)
  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: pEPQD0CM0705
FEATURES             Location/Qualifiers
     source          1..3945
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(87..181)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(202..861)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     776..795
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1023..1041
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1045..1088)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1091..1112
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             1129..2943
                     /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             2944..2964
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     terminator      3009..3066
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(3090..3107)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3261..3849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3341..3360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
       61 acttggtctg acagctcgag gcttggattc tcaccaataa aaaacgcccg gcggcaaccg
      121 agcgttctga acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt
      181 ccaagcgagc tcgatatcaa attacgcccc gccctgccac tcgtcgcagt actgttgtaa
      241 ttcattaagc attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc
      301 cagcggcatc agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc
      361 gaagaagttg tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt
      421 ggctgatacg aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc
      481 gtaacacgcc acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc
      541 actccagagt gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac
      601 actatcccat atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt
      661 catcaggcgg gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac
      721 ggtctttaaa aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac
      781 tgactgaaat gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata
      841 tccagtgatt tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa
      901 aaatacgccc ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccc
      961 gatcaactcg agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     1021 aaaataggcg tatcacgagg cagaatttca gataaaaaaa atccttagct ttcgctaagg
     1081 atgatttctg gaattcgcgg ccgcttctag agcgatgagt ggtctcgaat ggacacgtat
     1141 gccggcgctt atgatagaca atcaagagaa agagagaact cttctgccgc ctctccagct
     1201 acacagagat ctgccaacga ggataaagct gctgatttac aaagagaggt agagagagat
     1261 gggggtaggt ttagatttgt gggtcatttc tctgaggctc ctggtacatc agcctttggt
     1321 actgctgaac ggcctgaatt tgaaagaata ttaaatgagt gtcgggccgg tcgactaaat
     1381 atgataatag tttatgatgt atctaggttc agtagactca aagttatgga tgcaattccg
     1441 atcgtgagcg agctacttgc actcggtgtt actatcgtaa gtacacaaga aggtgtattc
     1501 aggcagggta atgtgatgga ccttatacac cttattatgc gactcgacgc tagccataaa
     1561 gagtcatcac tgaagagtgc taaaatcttg gacaccaaga acctgcaaag ggagcttgga
     1621 ggctatgtag gaggaaaagc tccgtatggt ttcgaactcg tgagtgagac aaaagagatt
     1681 accagaaatg gtaggatggt gaacgtggtt attaataagc tagctcatag cacaacccct
     1741 ctcacagggc cttttgagtt cgaacctgac gttattagat ggtggtggag agagatcaaa
     1801 actcacaaac atcttccatt taagccaggt agccaggctg ctatccaccc tggatctatc
     1861 actgggcttt gtaaaaggat ggatgctgac gcagtgccaa ctcgagggga aacaatcggt
     1921 aagaaaactg catcttccgc ttgggatccc gctacagtta tgagaatcct tcgggatccg
     1981 cgtattgctg gattcgccgc agaagtcatt tataagaaaa aacctgatgg tactcctact
     2041 acaaaaatcg aaggttatag gatccaaagg gatcctatta ctttgcgtcc agttgaatta
     2101 gactgtggac cgatcatcga acctgccgag tggtacgagt tgcaggcctg gcttgatggt
     2161 agagggcgtg gaaagggttt atctcgggga caggcaattt tgtcagcaat ggataagctt
     2221 tattgtgagt gtggggctgt tatgacatca aaacgtggag aagagtcaat caaggattct
     2281 tacaggtgca gacgtcgtaa agtggtggac ccttctgctc caggacagca tgagggtaca
     2341 tgcaacgttt ctatggctgc tcttgataaa tttgtggccg aacgtatttt caacaagatt
     2401 agacacgctg aaggggatga ggaaacttta gctcttcttt gggaggctgc ccggcgtttt
     2461 ggtaagctaa cagaggcacc agaaaaaagt ggcgaaagag ctaatttggt agccgaaaga
     2521 gcagacgcct tgaatgcatt ggaagaactt tacgaagata gggctgccgg agcatatgat
     2581 ggaccagttg gtcggaagca tttcaggaag caacaagctg cactaacttt gagacagcaa
     2641 ggcgcagagg agcgactcgc agagcttgag gccgccgagg ccccaaagct tccactcgat
     2701 caatggtttc ctgaggatgc cgacgctgat cctacaggtc caaagagctg gtggggaaga
     2761 gctagcgtag acgataagag agtgtttgtg ggtctatttg ttgacaagat tgtagttact
     2821 aagtccacta cgggtcgagg acagggcact cccatcgaaa agagagccag tattacctgg
     2881 gcaaagcctc caaccgatga tgacgaggac gatgcacaag atggtaccga ggatgtggca
     2941 gcaccaaaaa aaaagaggaa ggtgtaagct ttgagaccac tcatcgcgac tagtagcggc
     3001 cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct gccctttttc tttaaaaccg
     3061 aaaagattac ttcgcgttat gcaggcttcc tcgctcactg actcgctgcg ctcggtcgtt
     3121 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3181 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3241 aaggccgcgt tgctggcgtt tttccacagg ctccgccccc ctgacgagca tcacaaaaat
     3301 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3361 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3421 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3481 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3541 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3601 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3661 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3721 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3781 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3841 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3901 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttc
//
