LOCUS       sequence-223484-        7026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses glucocorticoid receptor in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7026)
  TITLE     Alice Wong Lab Unpublished Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7026)
  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            "Unpublished"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: pEGFP GR
FEATURES             Location/Qualifiers
     source          1..7026
                     /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"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1351..3684
                     /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"
     polyA_signal    3814..3935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3851..3870)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3905..3924
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3942..4397)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4079..4100)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4291..4310
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4424..4528
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4530..4887
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4530..4721
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4553..4573)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4738..4873
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4922..5716
                     /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             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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     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"
     primer_bind     complement(5904..5923)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5948..5995
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6324..6912
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6813..6832
                     /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 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct atggactcca aagaatcatt aactcctggt
     1381 agagaagaaa accccagcag tgtgcttgct caggagaggg gagatgtgat ggacttctat
     1441 aaaaccctaa gaggaggagc tactgtgaag gtttctgcgt cttcaccctc actggctgtc
     1501 gcttctcaat cagactccaa gcagcgaaga cttttggttg attttccaaa aggctcagta
     1561 agcaatgcgc agcagccaga tctgtccaaa gcagtttcac tctcaatggg actgtatatg
     1621 ggagagacag aaacaaaagt gatgggaaat gacctgggat tcccacagca gggccaaatc
     1681 agcctttcct cgggggaaac agacttaaag cttttggaag aaagcattgc aaacctcaat
     1741 aggtcgacca gtgttccaga gaaccccaag agttcagcat ccactgctgt gtctgctgcc
     1801 cccacagaga aggagtttcc aaaaactcac tctgatgtat cttcagaaca gcaacatttg
     1861 aagggccaga ctggcaccaa cggtggcaat gtgaaattgt ataccacaga ccaaagcacc
     1921 tttgacattt tgcaggattt ggagttttct tctgggtccc caggtaaaga gacgaatgag
     1981 agtccttgga gatcagacct gttgatagat gaaaactgtt tgctttctcc tctggcggga
     2041 gaagacgatt cattcctttt ggaaggaaac tcgaatgagg actgcaagcc tctcatttta
     2101 ccggacacta aacccaaaat taaggataat ggagatctgg ttttgtcaag ccccagtaat
     2161 gtaacactgc cccaagtgaa aacagaaaaa gaagatttca tcgaactctg cacccctggg
     2221 gtaattaagc aagagaaact gggcacagtt tactgtcagg caagctttcc tggagcaaat
     2281 ataattggta ataaaatgtc tgccatttct gttcatggtg tgagtacctc tggaggacag
     2341 atgtaccact atgacatgaa tacagcatcc ctttctcaac agcaggatca gaagcctatt
     2401 tttaatgtca ttccaccaat tcccgttggt tccgaaaatt ggaataggtg ccaaggatct
     2461 ggagatgaca acttgacttc tctggggact ctgaacttcc ctggtcgaac agttttttct
     2521 aatggctatt caagccccag catgagacca gatgtaagct ctcctccatc cagctcctca
     2581 acagcaacaa caggaccacc tcccaaactc tgcctggtgt gctctgatga agcttcagga
     2641 tgtcattatg gagtcttaac ttgtggaagc tgtaaagttt tcttcaaaag agcagtggaa
     2701 ggacagcaca attacctatg tgctggaagg aatgattgca tcatcgataa aattcgaaga
     2761 aaaaactgcc cagcatgccg ctatcgaaaa tgtcttcagg ctggaatgaa cctggaagct
     2821 cgaaaaacaa agaaaaaaat aaaaggaatt cagcaggcca ctacaggagt ctcacaagaa
     2881 acctctgaaa atcctggtaa caaaacaata gttcctgcaa cgttaccaca actcacccct
     2941 accctggtgt cactgttgga ggttattgaa cctgaagtgt tatatgcagg atatgatagc
     3001 tctgttccag actcaacttg gaggatcatg actacgctca acatgttagg agggcggcaa
     3061 gtgattgcag cagtgaaatg ggcaaaggca ataccaggtt tcaggaactt acacctggat
     3121 gaccaaatga ccctactgca gtactcctgg atgtttctta tggcatttgc tctggggtgg
     3181 agatcatata gacaatcaag tgcaaacctg ctgtgttttg ctcctgatct gattattaat
     3241 gagcagagaa tgactctacc ctgcatgtac gaccaatgta aacacatgct gtatgtttcc
     3301 tctgagttac acaggcttca ggtatcttat gaagagtatc tctgtatgaa aaccttactg
     3361 cttctctctt cagttcctaa ggacggtctg aagagccaag agctatttga tgaaattaga
     3421 atgacctaca tcaaagagct aggaaaagcc attgtcaaga gggaaggaaa ctccagccag
     3481 aactggcagc ggttttatca actgacaaaa ctcttggatt ctatgcatga agtggttgaa
     3541 aatctcctta actattgctt ccaaacattt ttggataaga ccatgagtat tgaattcccc
     3601 gagatgttag ctgaaatcat caccaatcag ataccaaaat attcaaatgg aaatatcaaa
     3661 aaacttctgt ttcatcaaaa gtgaggatcc accggatcta gataactgat cataatcagc
     3721 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     3781 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     3841 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     3901 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     3961 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4021 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4081 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4141 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4201 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4261 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4321 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4381 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4441 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4501 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4561 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4621 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4681 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4741 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4801 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4861 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     4921 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     4981 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5041 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5101 gcaagacgag 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 ctcaaggcga 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 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5821 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5881 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     5941 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     6001 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     6061 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     6121 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     6181 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6241 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6301 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6361 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6421 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6481 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6541 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6601 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6661 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6721 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6781 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6841 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6901 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6961 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7021 atgcat
//
