LOCUS       Exported                6610 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCI-nGL1-HEGO, complete sequence.
ACCESSION   AF061181
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6610)
  AUTHORS   Htun H, Holth LT, Walker D, Davie JR, Hager GL.
  TITLE     Direct visualization of the human estrogen receptor alpha reveals a 
            role for ligand in the nuclear distribution of the receptor
  JOURNAL   Mol. Biol. Cell 10 (2), 471-486 (1999)
  PUBMED    9950689
REFERENCE   2  (bases 1 to 6610)
  AUTHORS   Htun H, Holth L, Walker L, Davie JR, Hager GL.
  TITLE     Direct Submission
  JOURNAL   Submitted (22-APR-1998) Laboratory of Receptor Biology and Gene 
            Expression, National Cancer Institute, National Institutes of 
            Health, Bldg. 41/Rm. B517, 9000 Rockville Pike, Bethesda, MD 20892, 
            USA
REFERENCE   3  (bases 1 to 6610)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6610
                     /organism="Mammalian expression vector pCI-nGL1-HEGO"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:88432"
     regulatory      1..795
                     /regulatory_class="promoter"
                     /note="CMV IE enhancer/promoter region"
     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"
     intron          857..989
                     /note="chimeric"
     regulatory      1034..1052
                     /regulatory_class="promoter"
                     /note="T7"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1072..3672
                     /codon_start=1
                     /gene="GFP/ER fusion"
                     /product="green fluorescent protein-estrogen receptor alpha
                     fusion"
                     /label=GFP/ER fusion
                     /note="S65T variant of humanized green fluorescent protein 
                     fused to human estrogen receptor alpha"
                     /protein_id="AAD47137.1"
                     /translation="MAHHHHHHGYPYDVPDYAQSSAMSKGEELFTGVVPILVELDGDVN
                     GHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHD
                     FFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKL
                     EYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY
                     LSTQSALSKDPNEKRDHMVLLEFVTAAGITHGMDELYKGAGAGAGAGAISALTLHTKAS
                     GMALLHQIQGNELEPLNRPQLKIPLERPLGEVYLDSSKPAVYNYPEGAAYEFNAAAAAN
                     AQVYGQTGLPYGPGSEAAAFGSNGLGGFPPLNSVSPSPLMLLHPPPQLSPFLQPHGQQV
                     PYYLENEPSGYTVREAGPPAFYRPNSDNRRQGGRERLASTNDKGSMAMESAKETRYCAV
                     CNDYASGYHYGVWSCEGCKAFFKRSIQGHNDYMCPATNQCTIDKNRRKSCQACRLRKCY
                     EVGMMKGGIRKDRRGGRMLKHKRQRDDGEGRGEVGSAGDMRAANLWPSPLMIKRSKKNS
                     LALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRV
                     PGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVE
                     IFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKI
                     TDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLL
                     EMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSLQKYYITGEAEGFPATV"
     gene            1072..3672
                     /gene="GFP/ER fusion"
                     /label=GFP/ER fusion
     misc_feature    1078..1095
                     /gene="GFP/ER fusion"
                     /label=Region: polyhistidine tag
                     /note="Region: polyhistidine tag"
     CDS             1078..1095
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     misc_feature    1096..1125
                     /gene="GFP/ER fusion"
                     /note="recognized by monoclonal antibody 12CA5; Region: 
                     hemagglutinin tag (HA)"
     CDS             1099..1125
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     misc_feature    1126..1851
                     /gene="GFP/ER fusion"
                     /note="Region: S65T variant of humanized green fluorescent 
                     protein coding region"
     misc_feature    1852..1881
                     /gene="GFP/ER fusion"
                     /label=Region: glycine-alanine repeats
                     /note="Region: glycine-alanine repeats"
     misc_feature    1882..3669
                     /gene="GFP/ER fusion"
                     /note="Region: human estrogen receptor alpha coding region"
     CDS             2779..3672
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPVKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     regulatory      3713..3934
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 late polyadenylation signal"
     polyA_signal    3722..3843
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4024..4479
                     /label=Region: phage f1
                     /note="Region: phage f1"
     rep_origin      4024..4479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4813..4917
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4918..5778
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=bla
                     /protein_id="AAD47138.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            4918..5778
                     /gene="bla"
                     /label=bla
     rep_origin      5949..6537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(5982..6537)
                     /direction=LEFT
                     /label=ColE1 origin
                     /note="ColE1 origin"