LOCUS       sequence-492323-        5208 bp    DNA     circular SYN 07-NOV-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 5208)
  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 5208)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 7, 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: pEV824
FEATURES             Location/Qualifiers
     source          1..5208
                     /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             1921..2364
                     /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             2371..2391
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    2437..2461
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2491..2640
                     /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             2665..2685
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3115..3471
                     /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             3490..4059
                     /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    4100..4221
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4111..4130)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4165..4184
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      complement(4544..5132)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4624..4643)
                     /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 accggtctac catgggttga tcgctcgcaa ccatgggttg aggtgccttt
     1921 atggtgaagc tactgtcctc tatcgaacaa gcatgcgata tttgccgact taaaaagctc
     1981 aagtgctcca aagaaaaacc gaagtgcgcc aagtgtctga agaacaactg ggagtgtcgc
     2041 tactctccca aaaccaaaag gtccccgctg actagggcac atctgacaga agtggaatca
     2101 aggctagaaa gactggaaca gctatttcta ctgatttttc ctcgagagga ccttgacatg
     2161 attttgaaaa tggattcttt acaggatata aaagcattgt taacaggatt atttgtacaa
     2221 gataatgtga ataaagatgc cgtcacagat agattggctt cagtggagac tgatatgcct
     2281 ctaacattga gacagcatag aataagtgcg acatcatcat cggaagagag tagtaacaaa
     2341 ggtcaaagac agttgactgt atcgatgggc cccaagaaga agaggaaggt gggatctggc
     2401 tcgccaggga tccgtcgact tgacgcgttg atatcaacaa gtttgtacaa aaaagcaggc
     2461 tacaaagagg ccagcggttc cggacgggct gacgcattgg acgattttga tctggatatg
     2521 ctgggaagtg acgccctcga tgattttgac cttgacatgc ttggttcgga tgcccttgat
     2581 gactttgacc tcgacatgct cggcagtgac gcccttgatg atttcgacct ggacatgctg
     2641 attaactcta gaagttccgg atctccgaaa aagaaacgca aagttggtag ccagtacctg
     2701 cccgacaccg acgaccggca ccggatcgag gaaaagcgga agcggaccta cgagacattc
     2761 aagagcatca tgaagaagtc ccccttcagc ggccccaccg accctagacc tccacctaga
     2821 agaatcgccg tgcccagcag atccagcgcc agcgtgccaa aacctgcccc ccagccttac
     2881 cccttcacca gcagcctgag caccatcaac tacgacgagt tccctaccat ggtgttcccc
     2941 agcggccaga tctctcaggc ctctgctctg gctccagccc ctcctcaggt gctgcctcag
     3001 gctcctgctc ctgcaccagc tccagccatg gtgtctgcac tggctcaggc accagcaccc
     3061 gtgcctgtgc tggctcctgg acctccacag gctgtggctc caccagcccc taaacctaca
     3121 caggccggcg agggcacact gtctgaagct ctgctgcagc tgcagttcga cgacgaggat
     3181 ctgggagccc tgctgggaaa cagcaccgat cctgccgtgt tcaccgacct ggccagcgtg
     3241 gacaacagcg agttccagca gctgctgaac cagggcatcc ctgtggcccc tcacaccacc
     3301 gagcccatgc tgatggaata ccccgaggcc atcacccggc tcgtgacagg cgctcagagg
     3361 cctcctgatc cagctcctgc ccctctggga gcaccaggcc tgcctaatgg actgctgtct
     3421 ggcgacgagg acttcagctc tatcgccgat atggatttct cagccttgct gggctctggc
     3481 agcggcagcc gggattccag ggaagggatg tttttgccga agcctgaggc cggctccgct
     3541 attagtgacg tgtttgaggg ccgcgaggtg tgccagccaa aacgaatccg gccatttcat
     3601 cctccaggaa gtccatgggc caaccgccca ctccccgcca gcctcgcacc aacaccaacc
     3661 ggtccagtac atgagccagt cgggtcactg accccggcac cagtccctca gccactggat
     3721 ccagcgcccg cagtgactcc cgaggccagt cacctgttgg aggatcccga tgaagagacg
     3781 agccaggctg tcaaagccct tcgggagatg gccgatactg tgattcccca gaaggaagag
     3841 gctgcaatct gtggccaaat ggacctttcc catccgcccc caaggggcca tctggatgag
     3901 ctgacaacca cacttgagtc catgaccgag gatctgaacc tggactcacc cctgaccccg
     3961 gaattgaacg agattctgga taccttcctg aacgacgagt gcctcttgca tgccatgcat
     4021 atcagcacag gactgtccat cttcgacaca tctctgtttt gagcttccac tggattgtac
     4081 aattaccaac cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4141 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     4201 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     4261 ggggaggtgt gggaggtttt ttcgctcgct gaagagcggt gcgtttttat gcttgtagta
     4321 ttgtataatg tttttaagat ccttaattaa tagggataac agggtaatat cttgccctgc
     4381 acgaatacca gcgacccctt gcccaaatac ttgccgtggg cctcggcctg agagccaaaa
     4441 cacttgatgc ggaagaagtc ggtgcgctcc tgcttgtcgc cggcatcgtt gcgccacatc
     4501 taggatctgc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4561 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4621 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4681 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4741 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4801 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4861 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4921 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4981 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     5041 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     5101 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     5161 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatga
//
