LOCUS       sequence-459764-        4434 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 4434)
  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 4434)
  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-VPR
FEATURES             Location/Qualifiers
     source          1..4434
                     /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"
     protein_bind    1396..1420
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1450..1599
                     /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"
     CDS             1624..1644
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2074..2430
                     /codon_start=1
                     /product="transcriptional activation domain of human RelA, 
                     also known as p65 (O'Shea and Perkins, 2008)"
                     /label=RelA (p65) AD
                     /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
                     EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
                     EDFSSIADMDFSALL"
     CDS             2449..3018
                     /codon_start=1
                     /product="transcriptional activation domain from the human 
                     herpesvirus 4 (Epstein-Barr virus) replication and 
                     transcription activator Rta/BRLF1 (Hardwick et al., 1992; 
                     Chavez et al., 2015)"
                     /label=Rta AD
                     /translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
                     RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
                     REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
                     DTFLNDECLLHAMHISTGLSIFDTSLF"
     primer_bind     complement(3061..3079)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3147..3251
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3473..4264
                     /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 ggatcaggtt cgccagggat ccgtcgactt
     1381 gacgcgttga tatcaacaag tttgtacaaa aaagcaggct acaaagaggc cagcggttcc
     1441 ggacgggctg acgcattgga cgattttgat ctggatatgc tgggaagtga cgccctcgat
     1501 gattttgacc ttgacatgct tggttcggat gcccttgatg actttgacct cgacatgctc
     1561 ggcagtgacg cccttgatga tttcgacctg gacatgctga ttaactctag aagttccgga
     1621 tctccgaaaa agaaacgcaa agttggtagc cagtacctgc ccgacaccga cgaccggcac
     1681 cggatcgagg aaaagcggaa gcggacctac gagacattca agagcatcat gaagaagtcc
     1741 cccttcagcg gccccaccga ccctagacct ccacctagaa gaatcgccgt gcccagcaga
     1801 tccagcgcca gcgtgccaaa acctgccccc cagccttacc ccttcaccag cagcctgagc
     1861 accatcaact acgacgagtt ccctaccatg gtgttcccca gcggccagat ctctcaggcc
     1921 tctgctctgg ctccagcccc tcctcaggtg ctgcctcagg ctcctgctcc tgcaccagct
     1981 ccagccatgg tgtctgcact ggctcaggca ccagcacccg tgcctgtgct ggctcctgga
     2041 cctccacagg ctgtggctcc accagcccct aaacctacac aggccggcga gggcacactg
     2101 tctgaagctc tgctgcagct gcagttcgac gacgaggatc tgggagccct gctgggaaac
     2161 agcaccgatc ctgccgtgtt caccgacctg gccagcgtgg acaacagcga gttccagcag
     2221 ctgctgaacc agggcatccc tgtggcccct cacaccaccg agcccatgct gatggaatac
     2281 cccgaggcca tcacccggct cgtgacaggc gctcagaggc ctcctgatcc agctcctgcc
     2341 cctctgggag caccaggcct gcctaatgga ctgctgtctg gcgacgagga cttcagctct
     2401 atcgccgata tggatttctc agccttgctg ggctctggca gcggcagccg ggattccagg
     2461 gaagggatgt ttttgccgaa gcctgaggcc ggctccgcta ttagtgacgt gtttgagggc
     2521 cgcgaggtgt gccagccaaa acgaatccgg ccatttcatc ctccaggaag tccatgggcc
     2581 aaccgcccac tccccgccag cctcgcacca acaccaaccg gtccagtaca tgagccagtc
     2641 gggtcactga ccccggcacc agtccctcag ccactggatc cagcgcccgc agtgactccc
     2701 gaggccagtc acctgttgga ggatcccgat gaagagacga gccaggctgt caaagccctt
     2761 cgggagatgg ccgatactgt gattccccag aaggaagagg ctgcaatctg tggccaaatg
     2821 gacctttccc atccgccccc aaggggccat ctggatgagc tgacaaccac acttgagtcc
     2881 atgaccgagg atctgaacct ggactcaccc ctgaccccgg aattgaacga gattctggat
     2941 accttcctga acgacgagtg cctcttgcat gccatgcata tcagcacagg actgtccatc
     3001 ttcgacacat ctctgttttg agctttgaga ccacgaagtg gctcttcagt ggacgaaagg
     3061 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     3121 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3181 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3241 aaaggaagag tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta
     3301 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttttgggg tacagtctat
     3361 gcctcgggca tccaagcagc aagcgcgtta cgccgtgggt cgatgtttga tgttatggag
     3421 cagcaacgat gttacgcagc agggcagtcg ccctaaaaca aagttaaaca tcatgaggga
     3481 agcggtgatc gccgaagtat cgactcaact atcagaggta gttggcgtca tcgagcgcca
     3541 tctcgaaccg acgttgctgg ccgtacattt gtacggctcc gcagtggatg gcggcctgaa
     3601 gccacacagc gatattgatt tgctggttac ggtgaccgta aggcttgatg aaacaacgcg
     3661 gcgagctttg atcaacgacc ttttggaaac ttcggcttcc cctggagaga gcgagattct
     3721 ccgcgctgta gaagtcacca ttgttgtgca cgacgacatc attccgtggc gttatccagc
     3781 taagcgcgaa ctgcaatttg gagaatggca gcgcaatgac attcttgcag gtatcttcga
     3841 gccagccacg atcgacattg atctggctat cttgctgaca aaagcaagag aacatagcgt
     3901 tgccttggta ggtccagcgg cggaggaact ctttgatccg gttcctgaac aggatctatt
     3961 tgaggcgcta aatgaaacct taacgctatg gaactcgccg cccgactggg ctggcgatga
     4021 gcgaaatgta gtgcttacgt tgtcccgcat ttggtacagc gcagtaaccg gcaaaatcgc
     4081 gccgaaggat gtcgctgccg actgggcaat ggagcgcctg ccggcccagt atcagcccgt
     4141 catacttgaa gctagacagg cttatcttgg acaagaagaa gatcgcttgg cctcgcgcgc
     4201 agatcagttg gaagaatttg tccattacgt aaaaggcgag atcaccaagg tagtcggcaa
     4261 ataactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4321 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4381 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
