LOCUS       sequence-405574-        2467 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a kozak consensus sequence and a nucleus 
            signal peptide sequence with BC overhangs used to build level 1 
            constructs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2467)
  AUTHORS   Pollak B, Matute T, Nunez I, Cerda A, Lopez C, Vargas V, Kan A, 
            Bielinski V, von Dassow P, Dupont CL, Federici F
  TITLE     Universal loop assembly: open, efficient and cross-kingdom DNA 
            fabrication.
  JOURNAL   Synth Biol (Oxf). 2020;5(1):ysaa001. doi: 10.1093/synbio/ysaa001. 
            Epub 2020 Feb 5.
  PUBMED    32161816
REFERENCE   2  (bases 1 to 2467)
  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: "Synth Biol 
            (Oxf)."; date: "2020"; volume: "5(1):ysaa001. doi"; pages: " 
            10.1093/synbio/ysaa001. Epub 2020 Feb 5"
COMMENT     Sequence Label: pL0_BC-nuc_sig_pep
FEATURES             Location/Qualifiers
     source          1..2467
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1076..1085
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1457..1477
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     rep_origin      complement(1622..2210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1702..1721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2301..2467,1..625))
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg cgggacaacg
       61 taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat agcgttaagg
      121 tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt tcctccgccg
      181 ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata gccagatcaa
      241 tgtcgatcgt ggctggctcg aagataccag caagaatgtc attgcgctgc cattctccaa
      301 attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc acaacaatgg
      361 tgacttctac agcgcggaga atctcactct ctccagggga agccgaagtt tccaaaaggt
      421 cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta accagcaaat
      481 caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa tgtacggcca
      541 gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt tgagtcgata
      601 cttcggcgat caccgcttcc ctcatgcgaa acgatcctca tcctgtctct tgatcagata
      661 ttgatcccct gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg
      721 gcttcccaac cttaccagag ggcgccccag ctggcaattc cggttcgctt gctaagcttg
      781 catgcctgca agtcgactct agaggagcgg tgatcacagg cagcaacgct ctgtcatcgt
      841 tacaatcaac atgctaccct ccgcgagatc atccgtgttt caaacccggc agcttagttg
      901 ccgttcttcc gaatagcatc ggtaacatga gcaaagtctg ccgccttaca acggctctcc
      961 cgctgacgcc gtcccggact gatgggctgc ctgtatcgag tggtgatttt gtgccgagct
     1021 cattactcgc atccattctc aggctgtctc gtctcgtctc aggtctcata ctggggccac
     1081 catggataaa gttttaaaca gagaggaatc tttgcagcta atggaccttc taggtcttga
     1141 aaggagtgcc tgggggaata ttcctctgat gagaaaggca tatttaaaaa aatgcaagga
     1201 gtttcatcct gataaaggag gagatgaaga aaaaatgaag aaaatgaata ctctgtacaa
     1261 gaaaatggaa gatggagtaa aatatgctca tcaacctgac tttggaggct tctgggatgc
     1321 aactgagatt ccaacctatg gaactgatga atgggagcag tggtggaatg cctttaatga
     1381 ggaaaacctg ttttgctcag aagaaatgcc atctagtgat gatgaggcta ctgctgactc
     1441 tcaacattct actcctccaa aaaagaagag aaaggtacaa atgcgagacc accaggatac
     1501 atagattacc acaactccga gcccttccac ccacagaatc aggggataac gcaggaaaga
     1561 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1621 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1741 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1801 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1861 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1921 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1981 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2041 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2101 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2161 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2221 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2281 tcatgcattc taggtgatta ttatttgccg actaccttgg tgatctcgcc tttcacgtag
     2341 tggacaaatt cttccaactg atctgcgcgc gaggccaagc gatcttcttc ttgtccaaga
     2401 taagcctgtc tagcttcaag tatgacgggc tgatactggg ccggcaggcg ctccattgcc
     2461 cagtcgg
//
