LOCUS       sequence-254297-        7192 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses HA-tagged ER Y537S mutant in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7192)
  AUTHORS   Toy W, Shen Y, Won H, Green B, Sakr RA, Will M, Li Z, Gala K, 
            Fanning S, King TA, Hudis C, Chen D, Taran T, Hortobagyi G, Greene 
            G, Berger M, Baselga J, Chandarlapaty S
  TITLE     ESR1 ligand-binding domain mutations in hormone-resistant breast 
            cancer.
  JOURNAL   Nat Genet. 2013 Dec;45(12):1439-45. doi: 10.1038/ng.2822. Epub 2013 
            Nov 3.
  PUBMED    24185512
REFERENCE   2  (bases 1 to 7192)
  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"; doi: "10.1038/ng.2822"; 
            journalName: "Nat Genet"; date: "2013-12"; volume: "45"; issue: 
            "12"; pages: "1439-45"
COMMENT     Sequence Label: pcDNA-HA-ER Y537S
FEATURES             Location/Qualifiers
     source          1..7192
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(85..105)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        90..419
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      270..405
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     332..351
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             486..1280
                     /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(540..559)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1150..1169
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1454..1575
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1491..1510)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1545..1564
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1624..1640)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1624..1640)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1637..1659)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1648..1664
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1672..1678,1679..1696,1697..1702))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1717..1738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1855..1872)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2026..2614)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2106..2125)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2785..3576,3577..3645))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3408..3427
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3646..3750)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3825..3844)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4016..4395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4396..4599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4550..4570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4644..4663
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4644..4662
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             4725..4751
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             5595..6530
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLSDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             5643..6536
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLSDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     primer_bind     complement(6564..6581)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6570..6794
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(6840..7192,1..76)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6927..6946)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7137..7158
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
       61 aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
      121 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
      181 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      241 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
      301 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
      361 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
      421 agctcccggg agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt
      481 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
      541 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
      601 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
      661 aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
      721 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
      781 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
      841 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
      901 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
      961 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc
     1021 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     1081 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     1141 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     1201 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     1261 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     1321 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     1381 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     1441 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1501 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1561 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     1621 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     1681 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     1741 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     1801 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     1861 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     1921 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     1981 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2041 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2101 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2161 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2221 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2281 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     2341 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     2401 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     2461 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     2521 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     2581 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     2641 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     2701 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     2761 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     2821 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     2881 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     2941 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3001 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3061 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3121 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3181 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3241 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     3301 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     3361 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     3421 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     3481 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     3541 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     3601 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     3661 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     3721 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     3781 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     3841 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     3901 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     3961 cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat
     4021 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     4081 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4141 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4201 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4261 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4321 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4381 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     4441 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4501 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     4561 ggtaggcgtg tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc
     4621 actgcttact ggcttatcga aattaatacg actcactata gggagaccca agctggctag
     4681 cgtttaaact taagcttggt accgagctcg gatccgccac catgtatcca tatgacgtcc
     4741 cagactatgc caccatgacc ctccacacca aagcatctgg gatggcccta ctgcatcaga
     4801 tccaagggaa cgagctggag cccctgaacc gtccgcagct caagatcccc ctggagcggc
     4861 ccctgggcga ggtgtacctg gacagcagca agcccgccgt gtacaactac cccgagggcg
     4921 ccgcctacga gttcaacgcc gcggccgccg ccaacgcgca ggtctacggt cagaccggcc
     4981 tcccctacgg ccccgggtct gaggctgcgg cgttcggctc caacggcctg gggggtttcc
     5041 ccccactcaa cagcgtgtct ccgagcccgc tgatgctact gcacccgccg ccgcagctgt
     5101 cgcctttcct gcagccccac ggccagcagg tgccctacta cctggagaac gagcccagcg
     5161 gctacacggt gcgcgaggcc ggcccgccgg cattctacag gccaaattca gataatcgac
     5221 gccagggtgg cagagaaaga ttggccagta ccaatgacaa gggaagtatg gctatggaat
     5281 ctgccaagga gactcgctac tgtgcagtgt gcaatgacta tgcttcaggc taccattatg
     5341 gagtctggtc ctgtgagggc tgcaaggcct tcttcaagag aagtattcaa ggacataacg
     5401 actatatgtg tccagccacc aaccagtgca ccattgataa aaacaggagg aagagctgcc
     5461 aggcctgccg gctccgcaaa tgctacgaag tgggaatgat gaaaggtggg atacgaaaag
     5521 accgaagagg agggagaatg ttgaaacaca agcgccagag agatgatggg gagggcaggg
     5581 gtgaagtggg gtctgctgga gacatgagag ctgccaacct ttggccaagc ccgctcatga
     5641 tcaaacgctc taagaagaac agcctggcct tgtccctgac ggccgaccag atggtcagtg
     5701 ccttgttgga tgctgagccc cccatactct attccgagta tgatcctacc agacccttca
     5761 gtgaagcttc gatgatgggc ttactgacca acctggcaga cagggagctg gttcacatga
     5821 tcaactgggc gaagagggtg ccaggctttg tggatttgac cctccatgat caggtccacc
     5881 ttctagaatg tgcctggcta gagatcctga tgattggtct cgtctggcgc tccatggagc
     5941 acccagggaa gctactgttt gctcctaact tgctcttgga caggaaccag ggaaaatgtg
     6001 tagagggcat ggtggagatc ttcgacatgc tgctggctac atcatctcgg ttccgcatga
     6061 tgaatctgca gggagaggag tttgtgtgcc tcaaatctat tattttgctt aattctggag
     6121 tgtacacatt tctgtccagc accctgaagt ctctggaaga gaaggaccat atccaccgag
     6181 tcctggacaa gatcacagac actttgatcc acctgatggc caaggcaggc ctgaccctgc
     6241 agcagcagca ccagcggctg gcccagctcc tcctcatcct ctcccacatc aggcacatga
     6301 gtaacaaagg catggagcat ctgtacagca tgaagtgcaa gaacgtggtg cccctcagtg
     6361 acctgctgct ggagatgctg gacgcccacc gcctacatgc gcccactagc cgtggagggg
     6421 catccgtgga ggagacggac caaagccact tggccactgc gggctctact tcatcgcatt
     6481 ccttgcaaaa gtattacatc acgggggagg cagagggttt ccctgccacg gtctgattgg
     6541 gcccgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     6601 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     6661 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     6721 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     6781 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     6841 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     6901 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     6961 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     7021 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     7081 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     7141 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt tt
//
