LOCUS       40924_10711        6610 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian expression vector pCI-nGL1-HEGO, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6610)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol. Biol. 
            Cell"; date: "1999"; volume: "10"; issue: "2"; pages: "471-486"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "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"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6610
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        139..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
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1034..1052
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1078..1095
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             1099..1125
                     /codon_start=1
                     /label=HA
                     /note="HA (human influenza hemagglutinin) epitope tag"
                     /translation="YPYDVPDYA"
     CDS             1138..1851
                     /codon_start=1
                     /label=GFP (S65T)
                     /note="S65T variant of Aequorea victoria green fluorescent 
                     protein (Heim et al., 1995)"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     misc_feature    1852..1881
                     /gene="GFP/ER fusion"
                     /label=Region: glycine-alanine repeats
                     /note="Region: glycine-alanine repeats"
     CDS             2731..3666
                     /codon_start=1
                     /label=ERT2
                     /note="mutated ligand-binding domain of the human estrogen 
                     receptor (Feil et al., 1997)"
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     polyA_signal    complement(3722..3843)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      4024..4479
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4813..4917
                     /label=AmpR promoter
     CDS             4918..5775
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5949..6537
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 tcgtttagtg 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 acttaatacg actcactata ggctagcgaa ggagatccgc catggcccac
     1081 catcaccacc atcacggata tccatacgac gtgccagatt acgctcagtc gagtgccatg
     1141 agcaagggcg aggaactgtt cactggcgtg gtcccaattc tcgtggaact ggatggcgat
     1201 gtgaatgggc acaaattttc tgtcagcgga gagggtgaag gtgatgccac atacggaaag
     1261 ctcaccctga aattcatctg caccactgga aagctccctg tgccatggcc aacactggtc
     1321 actaccttca cctatggcgt gcagtgcttt tccagatacc cagaccatat gaagcagcat
     1381 gactttttca agagcgccat gcccgagggc tatgtgcagg agagaaccat ctttttcaaa
     1441 gatgacggga actacaagac ccgcgctgaa gtcaagttcg aaggtgacac cctggtgaat
     1501 agaatcgagc tgaagggcat tgactttaag gaggatggaa acattctcgg ccacaagctg
     1561 gaatacaact ataactccca caatgtgtac atcatggccg acaagcaaaa gaatggcatc
     1621 aaggtcaact tcaagatcag acacaacatt gaggatggat ccgtgcagct ggccgaccat
     1681 tatcaacaga acactccaat cggcgacggc cctgtgctcc tcccagacaa ccattacctg
     1741 tccacccagt ctgccctgtc taaagatccc aacgaaaaga gagaccacat ggtcctgctg
     1801 gagtttgtga ccgctgctgg gatcacacat ggcatggacg agctgtacaa gggcgccggc
     1861 gctggtgctg gtgctggcgc catcagcgcg ttgaccctcc acaccaaagc atctgggatg
     1921 gccctactgc atcagatcca agggaacgag ctggagcccc tgaaccgtcc gcagctcaag
     1981 atccccctgg agcggcccct gggcgaggtg tacctggaca gcagcaagcc cgccgtgtac
     2041 aactaccccg agggcgccgc ctacgagttc aacgccgcgg ccgccgccaa cgcgcaggtc
     2101 tacggtcaga ccggcctccc ctacggcccc gggtctgagg ctgcggcgtt cggctccaac
     2161 ggcctggggg gtttcccccc actcaacagc gtgtctccga gcccgctgat gctactgcac
     2221 ccgccgccgc agctgtcgcc tttcctgcag ccccacggcc agcaggtgcc ctactacctg
     2281 gagaacgagc ccagcggcta cacggtgcgc gaggccggcc cgccggcatt ctacaggcca
     2341 aattcagata atcgacgcca gggtggcaga gaaagattgg ccagtaccaa tgacaaggga
     2401 agtatggcta tggaatctgc caaggagact cgctactgtg cagtgtgcaa tgactatgct
     2461 tcaggctacc attatggagt ctggtcctgt gagggctgca aggccttctt caagagaagt
     2521 attcaaggac ataacgacta tatgtgtcca gccaccaacc agtgcaccat tgataaaaac
     2581 aggaggaaga gctgccaggc ctgccggctc cgcaaatgct acgaagtggg aatgatgaaa
     2641 ggtgggatac gaaaagaccg aagaggaggg agaatgttga aacacaagcg ccagagagat
     2701 gatggggagg gcaggggtga agtggggtct gctggagaca tgagagctgc caacctttgg
     2761 ccaagcccgc tcatgatcaa acgctctaag aagaacagcc tggccttgtc cctgacggcc
     2821 gaccagatgg tcagtgcctt gttggatgct gagcccccca tactctattc cgagtatgat
     2881 cctaccagac ccttcagtga agcttcgatg atgggcttac tgaccaacct ggcagacagg
     2941 gagctggttc acatgatcaa ctgggcgaag agggtgccag gctttgtgga tttgaccctc
     3001 catgatcagg tccaccttct agaatgtgcc tggctagaga tcctgatgat tggtctcgtc
     3061 tggcgctcca tggagcaccc agtgaagcta ctgtttgctc ctaacttgct cttggacagg
     3121 aaccagggaa aatgtgtaga gggcatggtg gagatcttcg acatgctgct ggctacatca
     3181 tctcggttcc gcatgatgaa tctgcaggga gaggagtttg tgtgcctcaa atctattatt
     3241 ttgcttaatt ctggagtgta cacatttctg tccagcaccc tgaagtctct ggaagagaag
     3301 gaccatatcc accgagtcct ggacaagatc acagacactt tgatccacct gatggccaag
     3361 gcaggcctga ccctgcagca gcagcaccag cggctggccc agctcctcct catcctctcc
     3421 cacatcaggc acatgagtaa caaaggcatg gagcatctgt acagcatgaa gtgcaagaac
     3481 gtggtgcccc tctatgacct gctgctggag atgctggacg cccaccgcct acatgcgccc
     3541 actagccgtg gaggggcatc cgtggaggag acggaccaaa gccacttggc cactgcgggc
     3601 tctacttcat cgcattcctt gcaaaagtat tacatcacgg gggaggcaga gggtttccct
     3661 gccacagtct gagagctccc tggcggaatt cggatcgggc ggccgcttcg agcagacatg
     3721 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     3781 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     3841 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggagat gtgggaggtt
     3901 ttttaaagca agtaaaacct ctacaaatgt ggtaaaatcg ataaggatcc gggctggcgt
     3961 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
     4021 tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4081 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4141 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4201 ttagagcttt acggcacctc gaccgcaaaa aacttgattt gggtgatggt tcacgtagtg
     4261 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4321 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4381 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaatat
     4441 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc gcctgatgcg gtattttctc
     4501 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     4561 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4621 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     4681 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     4741 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4801 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4861 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4921 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4981 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5041 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5101 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5161 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5221 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5281 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5341 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5401 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5461 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5521 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5581 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5641 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5701 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5761 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5821 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5881 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5941 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6001 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6061 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     6121 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6181 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6241 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6301 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6361 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6421 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6481 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6541 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatggct
     6601 cgacagatct
//
