LOCUS       sequence-492321-        5209 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 5209)
  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 5209)
  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: pEV823
FEATURES             Location/Qualifiers
     source          1..5209
                     /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
     CDS             1922..2365
                     /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             2372..2392
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    2438..2462
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2492..2641
                     /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             2666..2686
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3116..3472
                     /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             3491..4060
                     /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"
     polyA_signal    4101..4222
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4112..4131)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4166..4185
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4545..5133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4625..4644)
                     /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 accggtcctc ttatgggttg atcgctcgca accatgggtt gaggtgcctt
     1921 tatggtgaag ctactgtcct ctatcgaaca agcatgcgat atttgccgac ttaaaaagct
     1981 caagtgctcc aaagaaaaac cgaagtgcgc caagtgtctg aagaacaact gggagtgtcg
     2041 ctactctccc aaaaccaaaa ggtccccgct gactagggca catctgacag aagtggaatc
     2101 aaggctagaa agactggaac agctatttct actgattttt cctcgagagg accttgacat
     2161 gattttgaaa atggattctt tacaggatat aaaagcattg ttaacaggat tatttgtaca
     2221 agataatgtg aataaagatg ccgtcacaga tagattggct tcagtggaga ctgatatgcc
     2281 tctaacattg agacagcata gaataagtgc gacatcatca tcggaagaga gtagtaacaa
     2341 aggtcaaaga cagttgactg tatcgatggg ccccaagaag aagaggaagg tgggatctgg
     2401 ctcgccaggg atccgtcgac ttgacgcgtt gatatcaaca agtttgtaca aaaaagcagg
     2461 ctacaaagag gccagcggtt ccggacgggc tgacgcattg gacgattttg atctggatat
     2521 gctgggaagt gacgccctcg atgattttga ccttgacatg cttggttcgg atgcccttga
     2581 tgactttgac ctcgacatgc tcggcagtga cgcccttgat gatttcgacc tggacatgct
     2641 gattaactct agaagttccg gatctccgaa aaagaaacgc aaagttggta gccagtacct
     2701 gcccgacacc gacgaccggc accggatcga ggaaaagcgg aagcggacct acgagacatt
     2761 caagagcatc atgaagaagt cccccttcag cggccccacc gaccctagac ctccacctag
     2821 aagaatcgcc gtgcccagca gatccagcgc cagcgtgcca aaacctgccc cccagcctta
     2881 ccccttcacc agcagcctga gcaccatcaa ctacgacgag ttccctacca tggtgttccc
     2941 cagcggccag atctctcagg cctctgctct ggctccagcc cctcctcagg tgctgcctca
     3001 ggctcctgct cctgcaccag ctccagccat ggtgtctgca ctggctcagg caccagcacc
     3061 cgtgcctgtg ctggctcctg gacctccaca ggctgtggct ccaccagccc ctaaacctac
     3121 acaggccggc gagggcacac tgtctgaagc tctgctgcag ctgcagttcg acgacgagga
     3181 tctgggagcc ctgctgggaa acagcaccga tcctgccgtg ttcaccgacc tggccagcgt
     3241 ggacaacagc gagttccagc agctgctgaa ccagggcatc cctgtggccc ctcacaccac
     3301 cgagcccatg ctgatggaat accccgaggc catcacccgg ctcgtgacag gcgctcagag
     3361 gcctcctgat ccagctcctg cccctctggg agcaccaggc ctgcctaatg gactgctgtc
     3421 tggcgacgag gacttcagct ctatcgccga tatggatttc tcagccttgc tgggctctgg
     3481 cagcggcagc cgggattcca gggaagggat gtttttgccg aagcctgagg ccggctccgc
     3541 tattagtgac gtgtttgagg gccgcgaggt gtgccagcca aaacgaatcc ggccatttca
     3601 tcctccagga agtccatggg ccaaccgccc actccccgcc agcctcgcac caacaccaac
     3661 cggtccagta catgagccag tcgggtcact gaccccggca ccagtccctc agccactgga
     3721 tccagcgccc gcagtgactc ccgaggccag tcacctgttg gaggatcccg atgaagagac
     3781 gagccaggct gtcaaagccc ttcgggagat ggccgatact gtgattcccc agaaggaaga
     3841 ggctgcaatc tgtggccaaa tggacctttc ccatccgccc ccaaggggcc atctggatga
     3901 gctgacaacc acacttgagt ccatgaccga ggatctgaac ctggactcac ccctgacccc
     3961 ggaattgaac gagattctgg ataccttcct gaacgacgag tgcctcttgc atgccatgca
     4021 tatcagcaca ggactgtcca tcttcgacac atctctgttt tgagcttcca ctggattgta
     4081 caattaccaa ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat
     4141 gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat
     4201 tataagctgc aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca
     4261 gggggaggtg tgggaggttt tttcgctcgc tgaagagcgg tgcgttttta tgcttgtagt
     4321 attgtataat gtttttaaga tccttaatta atagggataa cagggtaata tcttgccctg
     4381 cacgaatacc agcgacccct tgcccaaata cttgccgtgg gcctcggcct gagagccaaa
     4441 acacttgatg cggaagaagt cggtgcgctc ctgcttgtcg ccggcatcgt tgcgccacat
     4501 ctaggatctg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4741 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4981 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5101 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     5161 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatga
//
