LOCUS       sequence-470668-        7107 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express GR-NanoLuc(R) Fusion Protein in Mammalian Cells under a CMV 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7107)
  AUTHORS   Promega
  TITLE     Promega Plasmid Collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7107)
  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: 
            "Unpublished"
COMMENT     Sequence Label: NanoBRET Assay Acceptor Vector, GR
FEATURES             Location/Qualifiers
     source          1..7107
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..721
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     672..692
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     intron          857..989
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     972..991
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     primer_bind     1033..1052
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1033..1051
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1065..3395
                     /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
                     LSQQQDQKPIFNVIPPIPVGSENWNRCQGSGDDNLTSLGTLNFPGRTVFSNGYSSPSMR
                     PDVSSPPSSSSTATTGPPPKLCLVCSDEASGCHYGVLTCGSCKVFFKRAVEGQHNYLCA
                     GRNDCIIDKIRRKNCPACRYRKCLQAGMNLEARKTKKKIKGIQQATTGVSQETSENPGN
                     KTIVPATLPQLTPTLVSLLEVIEPEVLYAGYDSSVPDSTWRIMTTLNMLGGRQVIAAVK
                     WAKAIPGFRNLHLDDQMTLLQYSWMFLMAFALGWRSYRQSSANLLCFAPDLIINEQRMT
                     LPCMYDQCKHMLYVSSELHRLQVSYEEYLCMKTLLLLSSVPKDGLKSQELFDEIRMTYI
                     KELGKAIVKREGNSSQNWQRFYQLTKLLDSMHEVVENLLNYCFQTFLDKTMSIEFPEML
                     AEIITNQIPKYSNGNIKKLLFHQK"
     CDS             3450..3962
                     /codon_start=1
                     /product="NanoLuc(R) luciferase"
                     /label=Nluc
                     /note="human codon-optimized"
                     /translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
                     VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
                     PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
                     LCERILA"
     primer_bind     complement(4050..4068)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     polyA_signal    4105..4226
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4116..4135)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4170..4189
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        4471..4828
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4471..4662
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4494..4514)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4679..4814
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        4843..4890
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4922..5716
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4976..4995)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5586..5605
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5780..5828
                     /note="synthetic polyadenylation signal"
     rep_origin      complement(6067..6655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6147..6166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    6771..7054
                     /label=cer region
                     /note="ColE1-derived recombination site that helps to 
                     maintain plasmids as monomers"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      661 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      721 tggtttagtg aaccgtcaga tcactagaag ctttattgcg gtagtttatc acagttaaat
      781 tgctaacgca gtcagtgctt ctgacacaac agtctcgaac ttaagctgca gaagttggtc
      841 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
      901 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
      961 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1021 aggctagagt attaatacga ctcactatag ggctagcgat cgccatggac tccaaagaat
     1081 cattaactcc tggtagagaa gaaaacccca gcagtgtgct tgctcaggag aggggagatg
     1141 tgatggactt ctataaaacc ctaagaggag gagctactgt gaaggtttct gcgtcttcac
     1201 cctcactggc tgtcgcttct caatcagact ccaagcagcg aagacttttg gttgattttc
     1261 caaaaggctc agtaagcaat gcgcagcagc cagatctgtc caaagcagtt tcactctcaa
     1321 tgggactgta tatgggagag acagaaacaa aagtgatggg aaatgacctg ggattcccac
     1381 agcagggcca aatcagcctt tcctcggggg aaacagactt aaagcttttg gaagaaagca
     1441 ttgcaaacct caataggtcg accagtgttc cagagaaccc caagagttca gcatccactg
     1501 ctgtgtctgc tgcccccaca gagaaggagt ttccaaaaac tcactctgat gtatcttcag
     1561 aacagcaaca tttgaagggc cagactggca ccaacggtgg caatgtgaaa ttgtatacca
     1621 cagaccaaag cacctttgac attttgcagg atttggagtt ttcttctggg tccccaggta
     1681 aagagacgaa tgagagtcct tggagatcag acctgttgat agatgaaaac tgtttgcttt
     1741 ctcctctggc gggagaagac gattcattcc ttttggaagg aaactcgaat gaggactgca
     1801 agcctctcat tttaccggac actaaaccca aaattaagga taatggagat ctggttttgt
     1861 caagccccag taatgtaaca ctgccccaag tgaaaacaga aaaagaagat ttcatcgaac
     1921 tctgcacccc tggggtaatt aagcaagaga aactgggcac agtttactgt caggcaagct
     1981 ttcctggagc aaatataatt ggtaataaaa tgtctgccat ttctgttcat ggtgtgagta
     2041 cctctggagg acagatgtac cactatgaca tgaatacagc atccctttct caacagcagg
     2101 atcagaagcc tatttttaat gtcattccac caattcccgt tggttccgaa aattggaata
     2161 ggtgccaagg atctggagat gacaacttga cttctctggg gactctgaac ttccctggtc
     2221 gaacagtttt ttctaatggc tattcaagcc ccagcatgag accagatgta agctctcctc
     2281 catccagctc ctcaacagca acaacaggac cacctcccaa actctgcctg gtgtgctctg
     2341 atgaagcttc aggatgtcat tatggagtct taacttgtgg aagctgtaaa gttttcttca
     2401 aaagagcagt ggaaggacag cacaattacc tatgtgctgg aaggaatgat tgcatcatcg
     2461 ataaaattcg aagaaaaaac tgcccagcat gccgctatcg aaaatgtctt caggctggaa
     2521 tgaacctgga agctcgaaaa acaaagaaaa aaataaaagg aattcagcag gccactacag
     2581 gagtctcaca agaaacctct gaaaatcctg gtaacaaaac aatagttcct gcaacgttac
     2641 cacaactcac ccctaccctg gtgtcactgt tggaggttat tgaacctgaa gtgttatatg
     2701 caggatatga tagctctgtt ccagactcaa cttggaggat catgactacg ctcaacatgt
     2761 taggagggcg gcaagtgatt gcagcagtga aatgggcaaa ggcaatacca ggtttcagga
     2821 acttacacct ggatgaccaa atgaccctac tgcagtactc ctggatgttt cttatggcat
     2881 ttgctctggg gtggagatca tatagacaat caagtgcaaa cctgctgtgt tttgctcctg
     2941 atctgattat taatgagcag agaatgactc taccctgcat gtacgaccaa tgtaaacaca
     3001 tgctgtatgt ttcctctgag ttacacaggc ttcaggtatc ttatgaagag tatctctgta
     3061 tgaaaacctt actgcttctc tcttcagttc ctaaggacgg tctgaagagc caagagctat
     3121 ttgatgaaat tagaatgacc tacatcaaag agctaggaaa agccattgtc aagagggaag
     3181 gaaactccag ccagaactgg cagcggtttt atcaactgac aaaactcttg gattctatgc
     3241 atgaagtggt tgaaaatctc cttaactatt gcttccaaac atttttggat aagaccatga
     3301 gtattgaatt ccccgagatg ttagctgaaa tcatcaccaa tcagatacca aaatattcaa
     3361 atggaaatat caaaaaactt ctgtttcatc aaaaggtttc tctcggtggt tcaggtggtg
     3421 gcgggagcgg tggagggagc agcggtggag tcttcacact cgaagatttc gttggggact
     3481 ggcgacagac agccggctac aacctggacc aagtccttga acagggaggt gtgtccagtt
     3541 tgtttcagaa tctcggggtg tccgtaactc cgatccaaag gattgtcctg agcggtgaaa
     3601 atgggctgaa gatcgacatc catgtcatca tcccgtatga aggtctgagc ggcgaccaaa
     3661 tgggccagat cgaaaaaatt tttaaggtgg tgtaccctgt ggatgatcat cactttaagg
     3721 tgatcctgca ctatggcaca ctggtaatcg acggggttac gccgaacatg atcgactatt
     3781 tcggacggcc gtatgaaggc atcgccgtgt tcgacggcaa aaagatcact gtaacaggga
     3841 ccctgtggaa cggcaacaaa attatcgacg agcgcctgat caaccccgac ggctccctgc
     3901 tgttccgagt aaccatcaac ggagtgaccg gctggcggct gtgcgaacgc attctggcgt
     3961 aaggccgcga ctctagagtc gacctgcagg catgcaagct gatccggctg ctaacaaagc
     4021 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
     4081 ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     4141 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     4201 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     4261 ttcaggggga gatgtgggag gtttttttaa gcaagtaaaa cctctacaaa tgtggtaaaa
     4321 tcgaatttta acaaaatatt aacgcttaca atttcctgat gcggtatttt ctccttacgc
     4381 atctgtgcgg tatttcacac cgcatacgcg gatctgcgca gcaccatggc ctgaaataac
     4441 ctctgaaaga ggaacttggt taggtacctt ctgaggcgga aagaaccagc tgtggaatgt
     4501 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     4561 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4621 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4681 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4741 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4801 cttttttgga ggcctaggct tttgcaaaaa gcttaattaa ctgttgacaa ttaatcatcg
     4861 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccca ggaggcagat
     4921 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4981 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5041 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5101 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5161 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5221 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5281 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5341 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5401 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5461 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5521 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5581 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5641 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5701 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5761 ctgccatcac gatggccgca ataaaatatc tttattttca ttacatctgt gtgttggttt
     5821 tttgtgtgaa tcgatagcga taaggatcct ctttgcgctt gcgttttccc ttgtccagat
     5881 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctaccc
     5941 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     6001 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     6061 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     6121 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     6181 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     6241 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     6301 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     6361 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     6421 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     6481 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     6541 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     6601 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
     6661 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     6721 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttatagt ccggaaatac
     6781 aggaacgcac gctggatggc ccttcgctgg gatggtgaaa ccatgaaaaa tggcagcttc
     6841 agtggattaa gtgggggtaa tgtggcctgt accctctggt tgcataggta ttcatacggt
     6901 taaaatttat caggcgcgat tgcggcagtt tttcgggtgg tttgttgcca tttttacctg
     6961 tctgctgccg tgatcgcgct gaacgcgttt tagcggtgcg tacaattaag ggattatggt
     7021 aaatccactt actgtctgcc ctcgtagcca tcgagataaa ccgcagtact ccggccacga
     7081 tgcgtccggc gtagaggatc gagatct
//
