LOCUS       sequence-322068-        4049 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  two repeats of ER (Estradiol receptor).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4049)
  TITLE     Orzaez lab plasmids
REFERENCE   2  (bases 1 to 4049)
  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"
COMMENT     Sequence Label: pUPD2 ER-ER (GB2666)
FEATURES             Location/Qualifiers
     source          1..4049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             55..990
                     /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             103..993
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     CDS             1024..1959
                     /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             1072..1962
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     terminator      2006..2063
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(2087..2104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2258..2846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2338..2357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3029..3123)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3144..3803)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3718..3737
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3965..3983
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3987..4030)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(4033..4049,1..5)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tagagcgatg agtggtctcg agccctgatt aacggcagcg gcagcggcag ctctgctgga
       61 gacatgagag ctgccaacct ttggccaagc ccgctcatga tcaaacgctc taagaagaac
      121 agcctggcct tgtccctgac ggccgaccag atggtcagtg ccttgttgga tgctgagccc
      181 cccatactct attccgagta tgatcctacc agacccttca gtgaagcttc gatgatgggc
      241 ttactgacca acctggcaga cagggagctg gttcacatga tcaactgggc gaagagggtg
      301 ccaggctttg tggatttgac cctccatgat caggtccacc ttctagaatg tgcctggcta
      361 gagatcctga tgattggact cgtctggcgc tccatggagc acccagggaa gctactgttt
      421 gctcctaact tgctcttgga caggaaccag ggaaaatgtg tagagggcat ggtggagatc
      481 ttcgacatgc tgctggctac atcatctcgg ttccgcatga tgaatctgca gggagaggag
      541 tttgtgtgcc tcaaatctat tattttgctt aattctggag tgtacacatt tctgtccagc
      601 accctgaagt ctctggaaga gaaggaccat atccaccgag tcctggacaa gatcacagac
      661 actttgatcc acctgatggc caaggcaggc ctgaccctgc agcagcagca ccagcggctg
      721 gcccagctcc tcctcatcct ctcccacatc aggcacatga gtaacaaagg catggagcat
      781 ctgtacagca tgaagtgcaa gaacgtggtg cccctctatg acctgctgct ggagatgctg
      841 gacgcccacc gcctacatgc gcccactagc cgtggagggg catccgtgga ggaaacggac
      901 caaagccact tggccactgc gggctctact tcgtcgcatt ccttgcaaaa gtattacatc
      961 acgggggagg cagagggttt ccctgccaca gtactgatta acggcagcgg cagcggcagc
     1021 tctgctggag acatgagagc tgccaacctt tggccaagcc cgctcatgat caaacgctct
     1081 aagaagaaca gcctggcctt gtccctgacg gccgaccaga tggtcagtgc cttgttggat
     1141 gctgagcccc ccatactcta ttccgagtat gatcctacca gacccttcag tgaagcttcg
     1201 atgatgggct tactgaccaa cctggcagac agggagctgg ttcacatgat caactgggcg
     1261 aagagggtgc caggctttgt ggatttgacc ctccatgatc aggtccacct tctagaatgt
     1321 gcctggctag agatcctgat gattggactc gtctggcgct ccatggagca cccagggaag
     1381 ctactgtttg ctcctaactt gctcttggac aggaaccagg gaaaatgtgt agagggcatg
     1441 gtggagatct tcgacatgct gctggctaca tcatctcggt tccgcatgat gaatctgcag
     1501 ggagaggagt ttgtgtgcct caaatctatt attttgctta attctggagt gtacacattt
     1561 ctgtccagca ccctgaagtc tctggaagag aaggaccata tccaccgagt cctggacaag
     1621 atcacagaca ctttgatcca cctgatggcc aaggcaggcc tgaccctgca gcagcagcac
     1681 cagcggctgg cccagctcct cctcatcctc tcccacatca ggcacatgag taacaaaggc
     1741 atggagcatc tgtacagcat gaagtgcaag aacgtggtgc ccctctatga cctgctgctg
     1801 gagatgctgg acgcccaccg cctacatgcg cccactagcc gtggaggggc atccgtggag
     1861 gaaacggacc aaagccactt ggccactgcg ggctctactt cgtcgcattc cttgcaaaag
     1921 tattacatca cgggggaggc agagggtttc cctgccacag taaattcgtg agaccactca
     1981 tcgcgactag tagcggccgc tgcagtccgg caaaaaaggg caaggtgtca ccaccctgcc
     2041 ctttttcttt aaaaccgaaa agattacttc gcgttatgca ggcttcctcg ctcactgact
     2101 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2161 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2221 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccacaggctc cgcccccctg
     2281 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2341 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2401 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2461 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2521 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2581 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2641 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2701 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2761 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2821 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2881 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2941 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3001 aaacttggtc tgacagctcg aggcttggat tctcaccaat aaaaaacgcc cggcggcaac
     3061 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     3121 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcgtcgca gtactgttgt
     3181 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     3241 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     3301 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     3361 ttggctgata cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     3421 ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     3481 tcactccaga gtgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     3541 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     3601 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     3661 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     3721 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     3781 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     3841 aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgtgc
     3901 ccgatcaact cgagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
     3961 taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa
     4021 ggatgatttc tggaattcgc ggccgcttc
//
