LOCUS       sequence-405585-        3868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 plasmid containing a beta glucuronidase reporter gene with 
            CD overhangs used to build a level 1 construct..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3868)
  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 3868)
  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_CD-GUS
FEATURES             Location/Qualifiers
     source          1..3868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             478..1269
                     /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"
     rep_origin      1360..1948
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1849..1868
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2093..3868,1..33))
                     /codon_start=1
                     /gene="uidA (or gusA) from E. coli"
                     /product="beta-glucuronidase"
                     /label=GUS
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
ORIGIN
        1 ttcacgggtt ggggtttcta caggacgtaa catttgagac ctgagacgag acgagacagc
       61 ctgagaatgg atgcgagtaa tgagctcggc acaaaatcac cactcgatac aggcagccca
      121 tcagtccggg acggcgtcag cgggagagcc gttgtaaggc ggcagacttt gctcatgtta
      181 ccgatgctat tcggaagaac ggcaactaag ctgccgggtt tgaaacacgg atgatctcgc
      241 ggagggtagc atgttgattg taacgatgac agagcgttgc tgcctgtgat caccgctcct
      301 ctagagtcga cttgcaggca tgcaagctta gcaagcgaac cggaattgcc agctggggcg
      361 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
      421 atctgatggc gcaggggatc aatatctgat caagagacag gatgaggatc gtttcgcatg
      481 agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg cgtcatcgag
      541 cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt ggatggcggc
      601 ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct tgatgaaaca
      661 acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg agagagtgag
      721 attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc gtggcgttat
      781 ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct tgctggtatc
      841 ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc aagagaacat
      901 agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc tgaacaggat
      961 ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga ctgggctggc
     1021 gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt aaccggcaaa
     1081 atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc ccagtatcag
     1141 cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg cttggcctcg
     1201 cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac caaggtagtc
     1261 ggcaaataat aatcacctag aatgcatgac caaaatccct taacgtgagt tttcgttcca
     1321 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     1381 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     1441 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     1501 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     1561 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     1621 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     1681 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     1741 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1801 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1861 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1921 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1981 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtggg
     2041 tggaagggct cggagttgtg gtaatctatg tatcctggtg gtctcgacct gattgtttgc
     2101 ctccctgctg cggtttttca ccgaagttca tgccagtcca gcgtttttgc agcagaaaag
     2161 ccgccgactt cggtttgcgg tcgcgagtga agatcccttt cttgttaccg ccaacgcgca
     2221 atatgccttg cgaggtcgca aaatcggcga aattccatac ctgttcaccg acgacggcgc
     2281 tgacgcgatc aaagacgcgg tgatacatat ccagccatgc acactgatac tcttcactcc
     2341 acatgtcggt gtacattgag tgcagcccgg ctaacgtatc cacgccgtat tcggtgatga
     2401 taatcggctg atgcagtttc tcctgccagg ccagaagttc tttttccagt accttctctg
     2461 ccgtttccaa atcgccgctt tggacatacc atccgtaata acggttcagg cacagcacat
     2521 caaagagatc gctgatggta tcggtgtgag cgtcgcagaa cattacattg acgcaggtga
     2581 tcggacgcgt cgggtcgagt ttacgcgttg cttccgccag tggcgcgaaa tattcccgtg
     2641 caccttgcgg acgggtatcc ggttcgttgg caatactcca catcaccacg cttgggtggt
     2701 ttttgtcacg cgctatcagc tctttaatcg cctgtaagtg cgcttgctga gtttccccgt
     2761 tgactgcctc ttcgctgtac agttctttcg gcttgttgcc cgcttcgaaa ccaatgccta
     2821 aagagaggtt aaagccgaca gcagcagttt catcaatcac cacgatgcca tgttcatctg
     2881 cccagtcgag catctcttca gcgtaagggt aatgcgaggt acggtaggag ttggccccaa
     2941 tccagtccat taatgcgtgg tcgtgcacca tcagcacgtt atcgaatcct ttgccacgca
     3001 agtccgcatc ttcatgacga ccaaagccag taaagtagaa cggtttgtgg ttaatcagga
     3061 actgttcgcc cttcactgcc actgaccgga tgccgacgcg aagcgggtag atatcacact
     3121 ctgtctggct tttggctgtg acgcacagtt catagagata accttcaccc ggttgccaga
     3181 ggtgcggatt caccacttgc aaagtcccgc tagtgccttg tccagttgca accacctgtt
     3241 gatccgcatc acgcagttca acgctgacat caccattggc caccacctgc cagtcaacag
     3301 acgcgtggtt acagtcttgc gcgacatgcg tcaccacggt gatatcgtcc acccaggtgt
     3361 tcggcgtggt gtagagcatt acgctgcgat ggattccggc atagttaaag aaatcatgga
     3421 agtaagactg ctttttcttg ccgttttcgt cggtaatcac cattcccggc gggatagtct
     3481 gccagttcag ttcgttgttc acacaaacgg tgatacgtac acttttcccg gcaataacat
     3541 acggcgtgac atcggcttca aatggcgtat agccgccctg atgctccatc acttcctgat
     3601 tattgaccca cactttgccg taatgagtga ccgcatcgaa acgcagcacg atacgctggc
     3661 ctgcccaacc tttcggtata aagacttcgc gctgatacca gacgttgccc gcataattac
     3721 gaatatctgc atcggcgaac tgatcgttaa aactgcctgg cacagcaatt gcccggcttt
     3781 cttgtaacgc gctttcccac caacgctgat caattccaca gttttcgcga tccagactga
     3841 atgcccacag gccgtcgagt tttttgat
//
