LOCUS       sequence-456027-        7015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Nrg1 protein for luciferase and barcode assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7015)
  AUTHORS   Popovic L, Brankatschk B, Palladino G, Rossner MJ, Wehr MC
  TITLE     Polypharmacological profiling across protein target families and 
            cellular pathways using the multiplexed cell-based assay platform 
            safetyProfiler reveals efficacy, potency and side effects of drugs.
  JOURNAL   Biomed Pharmacother. 2024 Nov;180:117523. doi: 
            10.1016/j.biopha.2024.117523. Epub 2024 Oct 14.
  PUBMED    39405910
REFERENCE   2  (bases 1 to 7015)
  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: "Biomed 
            Pharmacother."; date: "2024-11"; volume: "180:117523. doi"; pages: "
            10.1016/j.biopha.2024.117523. Epub 2024 Oct 14"
COMMENT     Sequence Label: pTag4C_Nrg1-GV-2HA
FEATURES             Location/Qualifiers
     source          1..7015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    632..656
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2598..2622)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2648..3088
                     /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             3131..3151
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3302..3535
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             3539..3565
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             3569..3595
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3803..3924
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3840..3859)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3894..3913
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3931..4386)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4068..4089)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4280..4299
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4413..4517
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4519..4876
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4519..4710
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4542..4562)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4727..4862
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4911..5705
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4911..5705
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4965..4984)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5575..5594
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5893..5912)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5937..5984
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6313..6901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6802..6821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttatc aacaagtttg tacaaaaaag caggctctac
      661 catgtctgag cgcaaagaag gcagaggcaa ggggaagggc aagaagaagg accggggatc
      721 ccgcgggaag cccgcgcccg ccgaaggcga cccgagccca gcattgcctc ccagattgaa
      781 agagatgaaa agccaggagt cagctgcagg ctccaagctc gtgcttcggt gtgaaaccag
      841 ctctgagtac tcctcactca gattcaaatg gttcaagaac gggaatgagc tgaaccgtag
      901 gaataaacca caaaacgtca agatacagaa gaagccaggg aagtcagagc ttcgaatcaa
      961 caaagcgtcc ctggctgact ctggagaata tatgtgcaaa gtgatcagca agttaggaaa
     1021 cgacagtgcc tctgccaaca tcaccattgt tgagtcaaac gacctcacca ctggcatgtc
     1081 agcctcaact gaaagaccct atgtgtcctc agagtctccc attagaatat cagtttcaac
     1141 agaaggcgca aatacttctt catccacatc tacatccacg actgggacca gccatctcat
     1201 aaagtgtgcg gagaaggaga aaactttctg tgtgaatgga ggcgagtgct tcatggtgaa
     1261 ggacctgtca aacccctcaa gatacttgtg caagtgccca aatgagttta ctggtgatcg
     1321 ttgccaaaac tacgtaatgg ccagcttcta caagcatctt gggattgaat ttatggaagc
     1381 ggaggagctc taccagaaga gggtactgac aattactggc atctgtatcg ccctgttggt
     1441 ggtcggcatc atgtgtgtgg tggcctactg caaaaccaag aaacagcggc agaagcttca
     1501 tgatcggctc cggcagagcc ttcggtcaga acgaaacaac atggtgaaca tagcgaatgg
     1561 ccctcaccat ccaaacccac caccagagaa tgtgcaactg gtgaatcaat atgtatctaa
     1621 aaacgtcatc tccagtgagc atattgtgga gagagaagtg gagacctcct tttccaccag
     1681 tcactacact tccacagctc atcactccac gactgtcacc cagactccta gtcacagctg
     1741 gagtaatggg cacacagaaa gcatcatttc agaaagccac tctgtaatca tgatgtcatc
     1801 ggtagagaac agcaggcaca gcagcccagc tgggggccca cgaggacgtc ttcatggcct
     1861 gggaggccct cgcgaatgta acagcttcct caggcatgcc agagaaaccc ctgactccta
     1921 cagagactct cctcatagtg aaaggtatgt atcagccatg accaccccgg ctcgtatgtc
     1981 acctgtagat ttccacacgc caagctcccc taaatcgccc ccttcggaaa tgtctccacc
     2041 cgtgtccagc atgacggtgt ccatgccctc tgtggcagtc agcccctttg tggaagaaga
     2101 gaggcctctg cttcttgtga cgccaccgag gctacgggag aagaagtatg atcatcaccc
     2161 ccagcaactc aactcctttc atcacaaccc tgcacatcag agtaccagcc tcccccctag
     2221 cccattgagg atagtggagg atgaggaata cgaaacgacc caggagtatg agccaattca
     2281 agagcctatt aagaaagtca ccaatagccg gcgggccaaa agaaccaagc ccaatggcca
     2341 cattgccaat aggttggaaa tggacagcaa cccaagttct gtgagcagta actcagaaag
     2401 tgagacagaa gatgaaagag taggtgaaga tacaccattc ctgggcatac agaaccccct
     2461 ggcagccagc cttgaggtgg cccctgcctt ccgtctggct gagagcagga ctaacccagc
     2521 aggccgcttc tccacacagg aagaattaca ggccaggctg tctagtgtaa tcgctaacca
     2581 agaccctatt gctgtagacc cagctttctt gtacaaagtg gttgatggta ccgatctggc
     2641 ccgggccatg aagctactgt cttctatcga acaagcatgc gatatttgcc gacttaaaaa
     2701 gctcaagtgc tccaaagaaa aaccgaagtg cgccaagtgt ctgaagaaca actgggagtg
     2761 tcgctactct cccaaaacca aaaggtctcc gctgactagg gcacatctga cagaagtgga
     2821 atcaaggcta gaaagactgg aacagctatt tctactgatt tttcctcgag aagaccttga
     2881 catgattttg aaaatggatt ctttacagga tataaaagca ttgttaacag gattatttgt
     2941 acaagataat gtgaataaag atgccgtcac agatagattg gcttcagtgg agactgatat
     3001 gcctctaaca ttgagacagc atagaataag tgcgacatca tcatcggaag agagtagtaa
     3061 caaaggtcaa agacagttga ctgtatcgcc ggaattcccg gggatcagct tgataattcg
     3121 attcaccatg cccaagaaga agaggaaggt ggcgtacagc cgcgcgcgta cgaaaaacaa
     3181 ttacgggtct accatcgagg gcctgctcga tctcccggac gacgacgccc ccgaagaggc
     3241 ggggctggcg gctccgcgcc tgtcctttct ccccgcggga cacacgcgca gactgtcgac
     3301 ggcccccccg accgatgtca gcctggggga cgagctccac ttagacggcg aggacgtcgc
     3361 gatggcgcat gccgacgcgc tagacgattt cgatctggac atgttggggg acggggattc
     3421 cccgggtccg ggatttaccc cccacgactc cgccccctac ggcgctctgg atatggccga
     3481 cttcgagttt gagcagatgt ttaccgatgc ccttggaatt gacgagtacg gtgggggcta
     3541 cccatacgat gttcctgact atgcgggcta tccctatgac gtcccggact atgcatgaac
     3601 tagtgaatta tcgaattctg cagccaagct aattccgggc gaatgtctta tgatttatga
     3661 tttttattat taacctgcag ggcggccgcg actctagatc ataatcagcc ataccacatt
     3721 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     3781 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     3841 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     3901 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     3961 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4021 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4081 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4141 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4201 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4261 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4321 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4381 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4441 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4501 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     4561 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4621 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4681 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4741 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4801 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4861 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     4921 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     4981 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     5041 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     5101 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     5161 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     5221 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     5281 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     5341 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     5401 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     5461 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     5521 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     5581 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     5641 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     5701 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     5761 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     5821 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     5881 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     5941 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     6001 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     6061 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     6121 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     6181 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6241 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6301 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6361 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6421 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     6481 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6541 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6601 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6661 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6721 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6781 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6841 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6901 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6961 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
