LOCUS       sequence-164290-        4578 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway Donor vector containing ESR1 , part of the Target 
            Accelerator Plasmid Collection..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4578)
  AUTHORS   Kim E, Ilic N, Shrestha Y, Zou L, Kamburov A, Zhu C, Yang X, Lubonja
            R, Tran N, Nguyen C, Lawrence MS, Piccioni F, Bagul M, Doench JG, 
            Chouinard CR, Wu X, Hogstrom L, Natoli T, Tamayo P, Horn H, Corsello
            SM, Lage K, Root DE, Subramanian A, Golub TR, Getz G, Boehm JS, Hahn
            WC
  TITLE     Systematic Functional Interrogation of Rare Cancer Variants 
            Identifies Oncogenic Alleles.
  JOURNAL   Cancer Discov. 2016 Jul;6(7):714-26. doi: 
            10.1158/2159-8290.CD-16-0160. Epub 2016 May 4.
  PUBMED    27147599
REFERENCE   2  (bases 1 to 4578)
  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: 
            "10.1158/2159-8290.CD-16-0160"; journalName: "Cancer Discov"; date: 
            "2016-07"; volume: "6"; issue: "7"; pages: "714-26"
COMMENT     Sequence Label: pDONR223_ESR1_WT
FEATURES             Location/Qualifiers
     source          1..4578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..65
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     57..74
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     58..74
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    90..189
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             1038..1973
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPGKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     CDS             1086..1976
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     protein_bind    complement(1978..2077)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2094..2113)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2095..2113)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2118..2134)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2118..2134)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2565..3356
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3449..4037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3938..3957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4191..4208
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(4367..4394)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(4486..4572)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcagttccc tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta
       61 aaacgacggc cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga
      121 cctgttcgtt gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtacaaaa
      181 aagttggcac catgaccatg accctccaca ccaaagcatc tgggatggcc ctactgcatc
      241 agatccaagg gaacgagctg gagcccctga accgtccgca gctcaagatc cccctggagc
      301 ggcccctggg cgaggtgtac ctggacagca gcaagcccgc cgtgtacaac taccccgagg
      361 gcgccgccta cgagttcaac gccgcggccg ccgccaacgc gcaggtctac ggtcagaccg
      421 gcctccccta cggccccggg tctgaggctg cggcgttcgg ctccaacggc ctggggggtt
      481 tccccccact caacagcgtg tctccgagcc cgctgatgct actgcacccg ccgccgcagc
      541 tgtcgccttt cctgcagccc cacggccagc aggtgcccta ctacctggag aacgagccca
      601 gcggctacac ggtgcgcgag gccggcccgc cggcattcta caggccaaat tcagataatc
      661 gacgccaggg tggcagagaa agattggcca gtaccaatga caagggaagt atggctatgg
      721 aatctgccaa ggagactcgc tactgtgcag tgtgcaatga ctatgcttca ggctaccatt
      781 atggagtctg gtcctgtgag ggctgcaagg ccttcttcaa gagaagtatt caaggacata
      841 acgactatat gtgtccagcc accaaccagt gcaccattga taaaaacagg aggaagagct
      901 gccaggcctg ccggctccgc aaatgctacg aagtgggaat gatgaaaggt gggatacgaa
      961 aagaccgaag aggagggaga atgttgaaac acaagcgcca gagagatgat ggggagggca
     1021 ggggtgaagt ggggtctgct ggagacatga gagctgccaa cctttggcca agcccgctca
     1081 tgatcaaacg ctctaagaag aacagcctgg ccttgtccct gacggccgac cagatggtca
     1141 gtgccttgtt ggatgctgag ccccccatac tctattccga gtatgatcct accagaccct
     1201 tcagtgaagc ttcgatgatg ggcttactga ccaacctggc agacagggag ctggttcaca
     1261 tgatcaactg ggcgaagagg gtgccaggct ttgtggattt gaccctccat gatcaggtcc
     1321 accttctaga atgtgcctgg ctagagatcc tgatgattgg tctcgtctgg cgctccatgg
     1381 agcacccagg gaagctactg tttgctccta acttgctctt ggacaggaac cagggaaaat
     1441 gtgtagaggg catggtggag atcttcgaca tgctgctggc tacatcatct cggttccgca
     1501 tgatgaatct gcagggagag gagtttgtgt gcctcaaatc tattattttg cttaattctg
     1561 gagtgtacac atttctgtcc agcaccctga agtctctgga agagaaggac catatccacc
     1621 gagtcctgga caagatcaca gacactttga tccacctgat ggccaaggca ggcctgaccc
     1681 tgcagcagca gcaccagcgg ctggcccagc tcctcctcat cctctcccac atcaggcaca
     1741 tgagtaacaa aggcatggag catctgtaca gcatgaagtg caagaacgtg gtgcccctct
     1801 atgacctgct gctggagatg ctggacgccc accgcctaca tgcgcccact agccgtggag
     1861 gggcatccgt ggaggagacg gaccaaagcc acttggccac tgcgggctct acttcatcgc
     1921 attccttgca aaagtattac atcacggggg aggcagaggg tttccctgcc acggtcttgc
     1981 caactttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     2041 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     2101 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     2161 tctgatgtta cattgcacaa gataaaaata tatcatcatg cctcctctag accagccagg
     2221 acagaaatgc ctcgacttcg ctgctaccca aggttgccgg gtgacgcaca ccgtggaaac
     2281 ggatgaaggc acgaacccag tggacataag cctgttcggt tcgtaagctg taatgcaagt
     2341 agcgtatgcg ctcacgcaac tggtccagaa ccttgaccga acgcagcggt ggtaacggcg
     2401 cagtggcggt tttcatggct tgttatgact gtttttttgg ggtacagtct atgcctcggg
     2461 catccaagca gcaagcgcgt tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg
     2521 atgttacgca gcagggcagt cgccctaaaa caaagttaaa cattatgagg gaagcggtga
     2581 tcgccgaagt atcgactcaa ctatcagagg tagttggcgt catcgagcgc catctcgaac
     2641 cgacgttgct ggccgtacat ttgtacggct ccgcagtgga tggcggcctg aagccacaca
     2701 gtgatattga tttgctggtt acggtgaccg taaggcttga tgaaacaacg cggcgagctt
     2761 tgatcaacga ccttttggaa acttcggctt cccctggaga gagcgagatt ctccgcgctg
     2821 tagaagtcac cattgttgtg cacgacgaca tcattccgtg gcgttatcca gctaagcgcg
     2881 aactgcaatt tggagaatgg cagcgcaatg acattcttgc aggtatcttc gagccagcca
     2941 cgatcgacat tgatctggct atcttgctga caaaagcaag agaacatagc gttgccttgg
     3001 taggtccagc ggcggaggaa ctctttgatc cggttcctga acaggatcta tttgaggcgc
     3061 taaatgaaac cttaacgcta tggaactcgc cgcccgactg ggctggcgat gagcgaaatg
     3121 tagtgcttac gttgtcccgc atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg
     3181 atgtcgctgc cgactgggca atggagcgcc tgccggccca gtatcagccc gtcatacttg
     3241 aagctagaca ggcttatctt ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt
     3301 tggaagaatt tgtccactac gtgaaaggcg agatcaccaa ggtagtcggc aaataaccct
     3361 cgagccaccc atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag
     3421 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     3481 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     3541 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     3601 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     3661 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     3721 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     3781 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     3841 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     3901 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     3961 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     4021 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     4081 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     4141 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     4201 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     4261 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     4321 caattaatac gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt
     4381 caggatggcc ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca
     4441 ccctccgggc cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag
     4501 gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt
     4561 tcgttttatt tgatgcct
//
