LOCUS       sequence-459758-        3009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lvl 0 5 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3009)
  AUTHORS   Duportet X, Wroblewska L, Guye P, Li Y, Eyquem J, Rieders J, 
            Rimchala T, Batt G, Weiss R
  TITLE     A platform for rapid prototyping of synthetic gene networks in 
            mammalian cells.
  JOURNAL   Nucleic Acids Res. 2014 Dec 1;42(21):13440-51. doi: 
            10.1093/nar/gku1082. Epub 2014 Nov 5.
  PUBMED    25378321
REFERENCE   2  (bases 1 to 3009)
  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"; doi: 
            "10.1093/nar/gku1082"; journalName: "Nucleic Acids Res"; date: 
            "2014-12-1- 1"; volume: "42"; issue: "21"; pages: "13440-51"
COMMENT     Sequence Label: lvl0-G-Gal4-VP16
FEATURES             Location/Qualifiers
     source          1..3009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             847..1287
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             1294..1314
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1324..1350
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1360..1593
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     primer_bind     complement(1636..1654)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1722..1826
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2048..2839
                     /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 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tcactctgtg gtctcaaggt
      841 gccaccatga agctactgtc ctctatcgaa caagcatgcg atatttgccg acttaaaaag
      901 ctcaagtgct ccaaagaaaa accgaagtgc gccaagtgtc tgaagaacaa ctgggagtgt
      961 cgctactctc ccaaaaccaa aaggtccccg ctgactaggg cacatctgac agaagtggaa
     1021 tcaaggctag aaagactgga acagctattt ctactgattt ttcctcgaga ggaccttgac
     1081 atgattttga aaatggattc tttacaggat ataaaagcat tgttaacagg attatttgta
     1141 caagataatg tgaataaaga tgccgtcaca gatagattgg cttcagtgga gactgatatg
     1201 cctctaacat tgagacagca tagaataagt gcgacatcat catcggaaga gagtagtaac
     1261 aaaggtcaaa gacagttgac tgtatcgatg ggccccaaga agaagaggaa ggtgggatct
     1321 ggctacccat acgacgtgcc agattacgcc ggatcaggtg cccccccgac cgatgtcagc
     1381 ctgggggacg agctccactt agacggcgag gacgtggcga tggcgcatgc cgacgcgcta
     1441 gacgatttcg atctggacat gttgggggac ggggattccc cgggtccggg atttaccccc
     1501 cacgactccg ccccctacgg cgctctggat atggccgact tcgagtttga gcagatgttt
     1561 accgatgccc ttggaattga cgagtacggt gggtgagctt tgagaccacg aagtggctct
     1621 tcagtggacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1681 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1741 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1801 ttcaataata ttgaaaaagg aagagtatgc gctcacgcaa ctggtccaga accttgaccg
     1861 aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc ttgttatgac tgtttttttt
     1921 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg
     1981 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt
     2041 aaacatcatg agggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg
     2101 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt
     2161 ggatggcggc ctgaagccac acagcgatat tgatttgctg gttacggtga ccgtaaggct
     2221 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg
     2281 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc
     2341 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct
     2401 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc
     2461 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc
     2521 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga
     2581 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt
     2641 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc
     2701 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg
     2761 cttggcctcg cgcgcagatc agttggaaga atttgtccat tacgtaaaag gcgagatcac
     2821 caaggtagtc ggcaaataac tgtcagacca agtttactca tatatacttt agattgattt
     2881 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2941 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3001 aggatcttc
//
