LOCUS       sequence-492296-        5133 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 5133)
  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 5133)
  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: pFC543
FEATURES             Location/Qualifiers
     source          1..5133
                     /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
     primer_bind     1920..1944
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             1992..2432
                     /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             2439..2459
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     protein_bind    2505..2529
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2559..2708
                     /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             2733..2753
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3183..3539
                     /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             3558..4127
                     /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    4159..4207
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(4469..5057)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4549..4568)
                     /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 cagtggggac aacttgtgcc gtatcagtcc cctccacccc acagtgggga
     1801 atgccatcag tcgtttggtc ccctccaccc cacagtgggg ttttacccta gtctacaagt
     1861 cccctccacc ccacagtggg gcgaggtagg cgtatacggt gggaggccta tataagcaga
     1921 gctcgtttag tgaaccgtca gatcgcctgg agaattccag ataagtttgt acaaaaaaga
     1981 gaggtgccac catgaagcta ctgtcctcta tcgaacaagc atgcgatatt tgccgactta
     2041 aaaagctcaa gtgctccaaa gaaaaaccga agtgcgccaa gtgtctgaag aacaactggg
     2101 agtgtcgcta ctctcccaaa accaaaaggt ccccgctgac tagggcacat ctgacagaag
     2161 tggaatcaag gctagaaaga ctggaacagc tatttctact gatttttcct cgagaggacc
     2221 ttgacatgat tttgaaaatg gattctttac aggatataaa agcattgtta acaggattat
     2281 ttgtacaaga taatgtgaat aaagatgccg tcacagatag attggcttca gtggagactg
     2341 atatgcctct aacattgaga cagcatagaa taagtgcgac atcatcatcg gaagagagta
     2401 gtaacaaagg tcaaagacag ttgactgtat cgatgggccc caagaagaag aggaaggtgg
     2461 gatctggctc gccagggatc cgtcgacttg acgcgttgat atcaacaagt ttgtacaaaa
     2521 aagcaggcta caaagaggcc agcggttccg gacgggctga cgcattggac gattttgatc
     2581 tggatatgct gggaagtgac gccctcgatg attttgacct tgacatgctt ggttcggatg
     2641 cccttgatga ctttgacctc gacatgctcg gcagtgacgc ccttgatgat ttcgacctgg
     2701 acatgctgat taactctaga agttccggat ctccgaaaaa gaaacgcaaa gttggtagcc
     2761 agtacctgcc cgacaccgac gaccggcacc ggatcgagga aaagcggaag cggacctacg
     2821 agacattcaa gagcatcatg aagaagtccc ccttcagcgg ccccaccgac cctagacctc
     2881 cacctagaag aatcgccgtg cccagcagat ccagcgccag cgtgccaaaa cctgcccccc
     2941 agccttaccc cttcaccagc agcctgagca ccatcaacta cgacgagttc cctaccatgg
     3001 tgttccccag cggccagatc tctcaggcct ctgctctggc tccagcccct cctcaggtgc
     3061 tgcctcaggc tcctgctcct gcaccagctc cagccatggt gtctgcactg gctcaggcac
     3121 cagcacccgt gcctgtgctg gctcctggac ctccacaggc tgtggctcca ccagccccta
     3181 aacctacaca ggccggcgag ggcacactgt ctgaagctct gctgcagctg cagttcgacg
     3241 acgaggatct gggagccctg ctgggaaaca gcaccgatcc tgccgtgttc accgacctgg
     3301 ccagcgtgga caacagcgag ttccagcagc tgctgaacca gggcatccct gtggcccctc
     3361 acaccaccga gcccatgctg atggaatacc ccgaggccat cacccggctc gtgacaggcg
     3421 ctcagaggcc tcctgatcca gctcctgccc ctctgggagc accaggcctg cctaatggac
     3481 tgctgtctgg cgacgaggac ttcagctcta tcgccgatat ggatttctca gccttgctgg
     3541 gctctggcag cggcagccgg gattccaggg aagggatgtt tttgccgaag cctgaggccg
     3601 gctccgctat tagtgacgtg tttgagggcc gcgaggtgtg ccagccaaaa cgaatccggc
     3661 catttcatcc tccaggaagt ccatgggcca accgcccact ccccgccagc ctcgcaccaa
     3721 caccaaccgg tccagtacat gagccagtcg ggtcactgac cccggcacca gtccctcagc
     3781 cactggatcc agcgcccgca gtgactcccg aggccagtca cctgttggag gatcccgatg
     3841 aagagacgag ccaggctgtc aaagcccttc gggagatggc cgatactgtg attccccaga
     3901 aggaagaggc tgcaatctgt ggccaaatgg acctttccca tccgccccca aggggccatc
     3961 tggatgagct gacaaccaca cttgagtcca tgaccgagga tctgaacctg gactcacccc
     4021 tgaccccgga attgaacgag attctggata ccttcctgaa cgacgagtgc ctcttgcatg
     4081 ccatgcatat cagcacagga ctgtccatct tcgacacatc tctgttttga gcttccactg
     4141 gattgtacaa ttaccaacaa taaaatatct ttattttcat tacatctgtg tgttggtttt
     4201 ttgtgtgcgc tcgctgaaga gcggtgcgtt tttatgcttg tagtattgta taatgttttt
     4261 aagatcctta attaataggg ataacagggt aatatcttgc cctgcacgaa taccagcgac
     4321 cccttgccca aatacttgcc gtgggcctcg gcctgagagc caaaacactt gatgcggaag
     4381 aagtcggtgc gctcctgctt gtcgccggca tcgttgcgcc acatctagga tctgccagga
     4441 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4501 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4561 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4621 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4681 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4741 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4801 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4861 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4921 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4981 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5041 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5101 acgaaaactc acgttaaggg attttggtca tga
//
