LOCUS       sequence-452418-        6706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ESR1 receptor overexpression for luciferase and barcode assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  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 6706)
  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: pBK_Gal4-DBD-ESR1
FEATURES             Location/Qualifiers
     source          1..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        21..125
                     /gene="bla"
                     /label=AmpR promoter
     promoter        127..484
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        127..318
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(150..170)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      335..470
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             519..1313
                     /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             519..1313
                     /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(573..592)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1183..1202
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1501..1520)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1545..1592
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1921..2509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2410..2429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2684..2987
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2988..3191
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3142..3162
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3226..3235
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3235..3264
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3274..3714
                     /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"
     protein_bind    3733..3757
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4609..5544
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             4657..5550
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     protein_bind    complement(5551..5575)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5713..5732)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5714..5732)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5739..5756)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5739..5755)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5748..5770)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    6117..6238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6154..6173)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6208..6227
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6245..6700)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6382..6403)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6594..6613
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
       61 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
      121 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
      181 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
      241 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
      301 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      361 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
      421 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      481 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
      541 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
      601 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
      661 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
      721 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
      781 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
      841 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
      901 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
      961 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     1021 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     1081 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     1141 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     1201 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     1261 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     1321 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     1381 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     1441 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     1501 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     1561 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     1621 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     1681 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     1741 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     1801 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1861 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1921 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1981 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2041 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2101 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2161 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2221 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2281 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2341 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2401 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2461 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2521 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2581 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
     2641 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     2701 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     2761 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     2821 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
     2881 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     2941 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     3001 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     3061 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     3121 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     3181 taagcagagc tggtttagtg aaccgtcaga tccgctagtg gatctgccac catggaacag
     3241 aagctgatct cagaagagga tctggcccgg gccatgaagc tactgtcttc tatcgaacaa
     3301 gcatgcgata tttgccgact taaaaagctc aagtgctcca aagaaaaacc gaagtgcgcc
     3361 aagtgtctga agaacaactg ggagtgtcgc tactctccca aaaccaaaag gtctccgctg
     3421 actagggcac atctgacaga agtggaatca aggctagaaa gactggaaca gctatttcta
     3481 ctgatttttc ctcgagaaga ccttgacatg attttgaaaa tggattcttt acaggatata
     3541 aaagcattgt taacaggatt atttgtacaa gataatgtga ataaagatgc cgtcacagat
     3601 agattggctt cagtggagac tgatatgcct ctaacattga gacagcatag aataagtgcg
     3661 acatcatcat cggaagagag tagtaacaaa ggtcaaagac agttgactgt atcgccggaa
     3721 ttcccgggga tcacaagttt gtacaaaaaa gcaggctcca ccatgaccat gaccctccac
     3781 accaaagcat ctgggatggc cctactgcat cagatccaag ggaacgagct ggagcccctg
     3841 aaccgtccgc agctcaagat ccccctggag cggcccctgg gcgaggtgta cctggacagc
     3901 agcaagcccg ccgtgtacaa ctaccccgag ggcgccgcct acgagttcaa cgccgcggcc
     3961 gccgccaacg cgcaggtcta cggtcagacc ggcctcccct acggccccgg gtctgaggct
     4021 gcggcgttcg gctccaacgg cctggggggt ttccccccac tcaacagcgt gtctccgagc
     4081 ccgctgatgc tactgcaccc gccgccgcag ctgtcgcctt tcctgcagcc ccacggccag
     4141 caggtgccct actacctgga gaacgagccc agcggctaca cggtgcgcga ggccggcccg
     4201 ccggcattct acaggccaaa ttcagataat cgacgccagg gtggcagaga aagattggcc
     4261 agtaccaatg acaagggaag tatggctatg gaatctgcca aggagactcg ctactgtgca
     4321 gtgtgcaatg actatgcttc aggctaccat tatggagtct ggtcctgtga gggctgcaag
     4381 gccttcttca agagaagtat tcaaggacat aacgactata tgtgtccagc caccaaccag
     4441 tgcaccattg ataaaaacag gaggaagagc tgccaggcct gccggctccg caaatgctac
     4501 gaagtgggaa tgatgaaagg tgggatacga aaagaccgaa gaggagggag aatgttgaaa
     4561 cacaagcgcc agagagatga tggggagggc aggggtgaag tggggtctgc tggagacatg
     4621 agagctgcca acctttggcc aagcccgctc atgatcaaac gctctaagaa gaacagcctg
     4681 gccttgtccc tgacggccga ccagatggtc agtgccttgt tggatgctga gccccccata
     4741 ctctattccg agtatgatcc taccagaccc ttcagtgaag cttcgatgat gggcttactg
     4801 accaacctgg cagacaggga gctggttcac atgatcaact gggcgaagag ggtgccaggc
     4861 tttgtggatt tgaccctcca tgatcaggtc caccttctag aatgtgcctg gctagagatc
     4921 ctgatgattg gtctcgtctg gcgctccatg gagcacccag ggaagctact gtttgctcct
     4981 aacttgctct tggacaggaa ccagggaaaa tgtgtagagg gcatggtgga gatcttcgac
     5041 atgctgctgg ctacatcatc tcggttccgc atgatgaatc tgcagggaga ggagtttgtg
     5101 tgcctcaaat ctattatttt gcttaattct ggagtgtaca catttctgtc cagcaccctg
     5161 aagtctctgg aagagaagga ccatatccac cgagtcctgg acaagatcac agacactttg
     5221 atccacctga tggccaaggc aggcctgacc ctgcagcagc agcaccagcg gctggcccag
     5281 ctcctcctca tcctctccca catcaggcac atgagtaaca aaggcatgga gcatctgtac
     5341 agcatgaagt gcaagaacgt ggtgcccctc tatgacctgc tgctggagat gctggacgcc
     5401 caccgcctac atgcgcccac tagccgtgga ggggcatccg tggaggagac ggaccaaagc
     5461 cacttggcca ctgcgggctc tacttcatcg cattccttgc aaaagtatta catcacgggg
     5521 gaggcagagg gtttccctgc cacagtctga acccagcttt cttgtacaaa gtggtgatcc
     5581 gtcgacctgc agccaagcta attccgggcg aatctcttat gatttatgat ttttattatt
     5641 aagcttctag agcggccgcg ggcccatcga ttttccaccc gggtggggta ccaggtaagt
     5701 gtacccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     5761 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     5821 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     5881 ctgaatggcg aatggagatc caatttttaa gtgtataatg tgttaaacta ctgattctaa
     5941 ttgtttgtgt attttagatt cacagtccca aggctcattt caggcccctc agtcctcaca
     6001 gtctgttcat gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa
     6061 acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact
     6121 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     6181 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac
     6241 gcgtaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     6301 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     6361 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     6421 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     6481 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     6541 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     6601 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     6661 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtg
//
