LOCUS       sequence-492305-        5217 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The plasmid comprises an indicible transcriptional unit encoding 
            Gal4-VPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5217)
  AUTHORS   Caliendo F, Vitu E, Wang J, Kuo SH, Sandt H, Enghuus CN, Tordoff J, 
            Estrada N, Collins JJ, Weiss R
  TITLE     Customizable gene sensing and response without altering endogenous 
            coding sequences.
  JOURNAL   Nat Chem Biol. 2024 Sep 12. doi: 10.1038/s41589-024-01733-y.
  PUBMED    39266721
REFERENCE   2  (bases 1 to 5217)
  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: "Nat Chem 
            Biol. 2024 Sep 12. doi: 10.1038/s41589-024-01733-y."
COMMENT     Sequence Label: pEV742
FEATURES             Location/Qualifiers
     source          1..5217
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(95..886,887..955))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     718..737
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(956..1060)
                     /gene="bla"
                     /label=AmpR promoter
     regulatory      1882..1891
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1888..2331
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MVKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKT
                     KRSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVN
                     KDAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     CDS             2338..2358
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    2404..2428
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2458..2607
                     /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             2632..2652
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3082..3438
                     /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             3457..4026
                     /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(4061..4078)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4067..4291
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(4553..5141)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4633..4652)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
       61 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      121 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      181 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagaac
      241 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      301 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      361 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      421 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      481 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      541 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      601 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
      661 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
      721 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
      781 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
      841 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
      901 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
      961 tcctttttca atattattga agcatttatc aaggttattg tctcatgagc ggatacatat
     1021 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacgaattgg ccagcgctgc
     1081 attaccctgt tatccctaac cggtttaccg agctcttatt ggttttcaaa cttcattgac
     1141 tgtgccgctc ttcaaccgga gttctgcaat tcatgtgatc tgcgcctggg agctcacggg
     1201 gacagccccc ccccaaagcc cccagggatg taattacgtc cctcccccgc tagggggcag
     1261 cagcgagccg cccggggctc cgctccggtc cggcgctccc cccgcatccc cgagccggca
     1321 gcgtgcgggg acagcccggg cacggggaag gtggcacggg atcgctttcc tctgaacgct
     1381 tctcgctgct ctttgagcct gcagacacct ggggggatac ggggaaaaag ctaggctgaa
     1441 agaggatctt agaatgacac gcgcctggga gctcacgggg acagcccccc cccaaagccc
     1501 ccagggatgt aattacgtcc ctcccccgct agggggcagc agcgagccgc ccggggctcc
     1561 gctccggtcc ggcgctcccc ccgcatcccc gagccggcag cgtgcgggga cagcccgggc
     1621 acggggaagg tggcacggga tcgctttcct ctgaacgctt ctcgctgctc tttgagcctg
     1681 cagacacctg gggggatacg gggaaaaagc tggatcataa ctcgacgtgc atactgtccc
     1741 ctccacccca cagtggggca tagcgccggc cgcgccgtcc cctccacccc acagtggggc
     1801 accatatgct ctagagggta tataatgggg gccaaatgct agtctactac cagagctcat
     1861 cgctagcgct accggtcagg tgccaccatg gtgaagctac tgtcctctat cgaacaagca
     1921 tgcgatattt gccgacttaa aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag
     1981 tgtctgaaga acaactggga gtgtcgctac tctcccaaaa ccaaaaggtc cccgctgact
     2041 agggcacatc tgacagaagt ggaatcaagg ctagaaagac tggaacagct atttctactg
     2101 atttttcctc gagaggacct tgacatgatt ttgaaaatgg attctttaca ggatataaaa
     2161 gcattgttaa caggattatt tgtacaagat aatgtgaata aagatgccgt cacagataga
     2221 ttggcttcag tggagactga tatgcctcta acattgagac agcatagaat aagtgcgaca
     2281 tcatcatcgg aagagagtag taacaaaggt caaagacagt tgactgtatc gatgggcccc
     2341 aagaagaaga ggaaggtggg atctggctcg ccagggatcc gtcgacttga cgcgttgata
     2401 tcaacaagtt tgtacaaaaa agcaggctac aaagaggcca gcggttccgg acgggctgac
     2461 gcattggacg attttgatct ggatatgctg ggaagtgacg ccctcgatga ttttgacctt
     2521 gacatgcttg gttcggatgc ccttgatgac tttgacctcg acatgctcgg cagtgacgcc
     2581 cttgatgatt tcgacctgga catgctgatt aactctagaa gttccggatc tccgaaaaag
     2641 aaacgcaaag ttggtagcca gtacctgccc gacaccgacg accggcaccg gatcgaggaa
     2701 aagcggaagc ggacctacga gacattcaag agcatcatga agaagtcccc cttcagcggc
     2761 cccaccgacc ctagacctcc acctagaaga atcgccgtgc ccagcagatc cagcgccagc
     2821 gtgccaaaac ctgcccccca gccttacccc ttcaccagca gcctgagcac catcaactac
     2881 gacgagttcc ctaccatggt gttccccagc ggccagatct ctcaggcctc tgctctggct
     2941 ccagcccctc ctcaggtgct gcctcaggct cctgctcctg caccagctcc agccatggtg
     3001 tctgcactgg ctcaggcacc agcacccgtg cctgtgctgg ctcctggacc tccacaggct
     3061 gtggctccac cagcccctaa acctacacag gccggcgagg gcacactgtc tgaagctctg
     3121 ctgcagctgc agttcgacga cgaggatctg ggagccctgc tgggaaacag caccgatcct
     3181 gccgtgttca ccgacctggc cagcgtggac aacagcgagt tccagcagct gctgaaccag
     3241 ggcatccctg tggcccctca caccaccgag cccatgctga tggaataccc cgaggccatc
     3301 acccggctcg tgacaggcgc tcagaggcct cctgatccag ctcctgcccc tctgggagca
     3361 ccaggcctgc ctaatggact gctgtctggc gacgaggact tcagctctat cgccgatatg
     3421 gatttctcag ccttgctggg ctctggcagc ggcagccggg attccaggga agggatgttt
     3481 ttgccgaagc ctgaggccgg ctccgctatt agtgacgtgt ttgagggccg cgaggtgtgc
     3541 cagccaaaac gaatccggcc atttcatcct ccaggaagtc catgggccaa ccgcccactc
     3601 cccgccagcc tcgcaccaac accaaccggt ccagtacatg agccagtcgg gtcactgacc
     3661 ccggcaccag tccctcagcc actggatcca gcgcccgcag tgactcccga ggccagtcac
     3721 ctgttggagg atcccgatga agagacgagc caggctgtca aagcccttcg ggagatggcc
     3781 gatactgtga ttccccagaa ggaagaggct gcaatctgtg gccaaatgga cctttcccat
     3841 ccgcccccaa ggggccatct ggatgagctg acaaccacac ttgagtccat gaccgaggat
     3901 ctgaacctgg actcacccct gaccccggaa ttgaacgaga ttctggatac cttcctgaac
     3961 gacgagtgcc tcttgcatgc catgcatatc agcacaggac tgtccatctt cgacacatct
     4021 ctgttttgag cttccactgg attgtacaat taccaaccag cctcgactgt gccttctagt
     4081 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     4141 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     4201 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     4261 aggcatgctg gggatgcggt gggctctatg gcgctcgctg aagagcggtg cgtttttatg
     4321 cttgtagtat tgtataatgt ttttaagatc cttaattaat agggataaca gggtaatatc
     4381 ttgccctgca cgaataccag cgaccccttg cccaaatact tgccgtgggc ctcggcctga
     4441 gagccaaaac acttgatgcg gaagaagtcg gtgcgctcct gcttgtcgcc ggcatcgttg
     4501 cgccacatct aggatctgcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4561 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4621 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4681 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     4741 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     4801 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     4861 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     4921 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     4981 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5041 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5101 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5161 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatga
//
