LOCUS       sequence-459747-        2961 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 2961)
  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 2961)
  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-VP64
FEATURES             Location/Qualifiers
     source          1..2961
                     /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             1384..1533
                     /codon_start=1
                     /product="tetrameric repeat of the minimal activation 
                     domain of herpes simplex virus VP16 (Beerli et al., 1998)"
                     /label=VP64
                     /translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
                     DLDML"
     primer_bind     complement(1588..1606)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1674..1778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2000..2791
                     /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 aggccagcgg ttccggacgg
     1381 gctgacgcat tggacgattt tgatctggat atgctgggaa gtgacgccct cgatgatttt
     1441 gaccttgaca tgcttggttc ggatgccctt gatgactttg acctcgacat gctcggcagt
     1501 gacgcccttg atgatttcga cctggacatg ctgattaact ctagatgagc tttgagacca
     1561 cgaagtggct cttcagtgga cgaaagggcc tcgtgatacg cctattttta taggttaatg
     1621 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     1681 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1741 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gcgctcacgc aactggtcca
     1801 gaaccttgac cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg
     1861 actgtttttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     1921 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     1981 taaaacaaag ttaaacatca tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     2041 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     2101 cggctccgca gtggatggcg gcctgaagcc acacagcgat attgatttgc tggttacggt
     2161 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     2221 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     2281 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     2341 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     2401 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     2461 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     2521 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     2581 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     2641 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     2701 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc attacgtaaa
     2761 aggcgagatc accaaggtag tcggcaaata actgtcagac caagtttact catatatact
     2821 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2881 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2941 agaaaagatc aaaggatctt c
//
