LOCUS       sequence-456017-        7253 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GR receptor overexpression for luciferase and barcode assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7253)
  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 7253)
  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-GR(NR3C1)
FEATURES             Location/Qualifiers
     source          1..7253
                     /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             3763..6096
                     /codon_start=1
                     /gene="Homo sapiens NR3C1"
                     /product="human glucocorticoid receptor, isoform alpha"
                     /label=glucocorticoid receptor
                     /note="acts as a transcription factor that binds to 
                     glucocorticoid response elements, and as a modulator of 
                     other transcription factors"
                     /translation="MDSKESLTPGREENPSSVLAQERGDVMDFYKTLRGGATVKVSASS
                     PSLAVASQSDSKQRRLLVDFPKGSVSNAQQPDLSKAVSLSMGLYMGETETKVMGNDLGF
                     PQQGQISLSSGETDLKLLEESIANLNRSTSVPENPKSSASTAVSAAPTEKEFPKTHSDV
                     SSEQQHLKGQTGTNGGNVKLYTTDQSTFDILQDLEFSSGSPGKETNESPWRSDLLIDEN
                     CLLSPLAGEDDSFLLEGNSNEDCKPLILPDTKPKIKDNGDLVLSSPSNVTLPQVKTEKE
                     DFIELCTPGVIKQEKLGTVYCQASFPGANIIGNKMSAISVHGVSTSGGQMYHYDMNTAS
                     LSQQQDQKPILNVIPPIPVGSENWNRCQGSGDDNLTSLGTLNFPGRTVFSNGYSSPSMR
                     PDVSSPPSSSSTATTGPPPKLCLVCSDEASGCHYGVLTCGSCKVFFKRAVEGQHNYLCA
                     GRNDCIIDKIRRKNCPACRYRKCLQAGMNLGARKTKKKIKGIQQATTGVSQETSENPGN
                     KTIVPATLPQLTPTLVSLLEVIEPEVLYAGYDSSVPDSTWRIMTTLNMLGGRQVIAAVK
                     WAKAIPGFRNLHLDDQMTLLQYSRMFLMAFALGWRSYRQPSANLLCFAPDLIINEQRMT
                     LPCMYDQCKHMLYVSSELHRLQVSYEEYLCMKTLLLLSSVPKDGLKSQELFDEIRMTYI
                     KELGKAIVKREGNSSQNWQRFYQLTKLLDSMHEVVENLLNYCFQTFLDKTMSIEFPEML
                     AEIITNQIPKYSNGNIKKLLFHQK"
     protein_bind    complement(6098..6122)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(6260..6279)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6261..6279)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6286..6303)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6286..6302)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6295..6317)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    6664..6785
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6701..6720)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6755..6774
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6792..7247)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6929..6950)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7141..7160
                     /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 ccatggactc caaagaatca
     3781 ttaactcctg gtagagaaga aaaccccagc agtgtgcttg ctcaggagag gggagatgtg
     3841 atggacttct ataaaaccct aagaggagga gctactgtga aggtttctgc gtcttcaccc
     3901 tcactggctg tcgcttctca atcagactcc aagcagcgaa gacttttggt tgattttcca
     3961 aaaggctcag taagcaatgc gcagcagcca gatctgtcca aagcagtttc actctcaatg
     4021 ggactgtata tgggagagac agaaacaaaa gtgatgggaa atgacctggg attcccacag
     4081 cagggccaaa tcagcctttc ctcgggggaa acagacttaa agcttttgga agaaagcatt
     4141 gcaaacctca ataggtcgac cagtgttcca gagaacccca agagttcagc atccactgct
     4201 gtgtctgctg cccccacaga gaaggagttt ccaaaaactc actctgatgt atcttcagaa
     4261 cagcaacatt tgaagggcca gactggcacc aacggtggca atgtgaaatt gtataccaca
     4321 gaccaaagca cctttgacat tttgcaggat ttggagtttt cttctgggtc cccaggtaaa
     4381 gagacgaatg agagtccttg gagatcagac ctgttgatag atgaaaactg tttgctttct
     4441 cctctggcgg gagaagacga ttcattcctt ttggaaggaa actcgaatga ggactgcaag
     4501 cctctcattt taccggacac taaacccaaa attaaggata atggagatct ggttttgtca
     4561 agccccagta atgtaacact gccccaagtg aaaacagaaa aagaagattt catcgaactc
     4621 tgcacccctg gggtaattaa gcaagagaaa ctgggcacag tttactgtca ggcaagcttt
     4681 cctggagcaa atataattgg taataaaatg tctgccattt ctgttcatgg tgtgagtacc
     4741 tctggaggac agatgtacca ctatgacatg aatacagcat ccctttctca acagcaggat
     4801 cagaagccta ttcttaatgt cattccacca attcccgttg gttccgaaaa ttggaatagg
     4861 tgccaaggat ctggagatga caacttgact tctctgggga ctctgaactt ccctggtcga
     4921 acagtttttt ctaatggcta ttcaagcccc agcatgagac cagatgtaag ctctcctcca
     4981 tccagctcct caacagcaac aacaggacca cctcccaaac tctgcctggt gtgctctgat
     5041 gaagcttcag gatgtcatta tggagtctta acttgtggaa gctgtaaagt tttcttcaaa
     5101 agagcagtgg aaggacagca caattaccta tgtgctggaa ggaatgattg catcatcgat
     5161 aaaattcgaa gaaaaaactg cccagcatgc cgctatcgaa aatgtcttca ggctggaatg
     5221 aacctgggag ctcgaaaaac aaagaaaaaa ataaaaggaa ttcagcaggc cactacagga
     5281 gtctcacaag aaacctctga aaatcctggt aacaaaacaa tagttcctgc aacgttacca
     5341 caactcaccc ctaccctggt gtcactgttg gaggttattg aacctgaagt gttatatgca
     5401 ggatatgata gctctgttcc agactcaact tggaggatca tgactacgct caacatgtta
     5461 ggagggcggc aagtgattgc agcagtgaaa tgggcaaagg caataccagg tttcaggaac
     5521 ttacacctgg atgaccaaat gaccctactg cagtactcca ggatgtttct tatggcattt
     5581 gctctggggt ggagatcata tagacaacca agtgcaaacc tgctgtgttt tgctcctgat
     5641 ctgattatta atgagcagag aatgactcta ccctgcatgt acgaccaatg taaacacatg
     5701 ctgtatgttt cctctgagtt acacaggctt caggtatctt atgaagagta tctctgtatg
     5761 aaaaccttac tgcttctctc ttcagttcct aaggacggtc tgaagagcca agagctattt
     5821 gatgaaatta gaatgaccta catcaaagag ctaggaaaag ccattgtcaa gagggaagga
     5881 aactccagcc agaactggca gcggttttat caactgacaa aactcttgga ttctatgcat
     5941 gaagtggttg aaaatctcct taactattgc ttccaaacat ttttggataa gaccatgagt
     6001 attgaattcc ccgagatgtt agctgaaatc atcaccaatc agataccaaa atactcaaac
     6061 ggaaatatca aaaaacttct gtttcatcaa aagtgagacc cagctttctt gtacaaagtg
     6121 gtgatccgtc gacctgcagc caagctaatt ccgggcgaat ctcttatgat ttatgatttt
     6181 tattattaag cttctagagc ggccgcgggc ccatcgattt tccacccggg tggggtacca
     6241 ggtaagtgta cccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt
     6301 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     6361 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     6421 gcgcagcctg aatggcgaat ggagatccaa tttttaagtg tataatgtgt taaactactg
     6481 attctaattg tttgtgtatt ttagattcac agtcccaagg ctcatttcag gcccctcagt
     6541 cctcacagtc tgttcatgat cataatcagc cataccacat ttgtagaggt tttacttgct
     6601 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     6661 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     6721 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     6781 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     6841 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     6901 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     6961 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     7021 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     7081 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     7141 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     7201 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtg
//
