LOCUS       sequence-162852-        5077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5077)
  AUTHORS   Chang C, Norris JD, Gron H, Paige LA, Hamilton PT, Kenan DJ, Fowlkes
            D, McDonnell DP
  TITLE     Dissection of the LXXLL nuclear receptor-coactivator interaction 
            motif using combinatorial peptide libraries: discovery of peptide 
            antagonists of estrogen receptors alpha and beta.
  JOURNAL   Mol Cell Biol. 1999 Dec . 19(12):8226-39.
  PUBMED    10567548
REFERENCE   2  (bases 1 to 5077)
  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"; journalName: "Mol Cell 
            Biol. 1999 Dec . 19(12):8226-39."
COMMENT     Sequence Label: VP16-ER alpha
FEATURES             Location/Qualifiers
     source          1..5077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(30..52)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     152..171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        252..270
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     252..269
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(280..300)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     regulatory      608..617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             620..640
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             641..874
                     /codon_start=1
                     /gene="UL48"
                     /product="transcriptional activation domain of herpes 
                     simplex virus protein VP16 (Triezenberg et al., 1988; 
                     Cousens et al., 1989)"
                     /label=VP16 AD
                     /translation="APPTDVSLGDELHLDGEDVAMAHADALDDFDLDMLGDGDSPGPGF
                     TPHDSAPYGALDMADFEFEQMFTDALGIDEYGG"
     CDS             1727..2662
                     /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             1775..2668
                     /codon_start=1
                     /gene="ESR1"
                     /product="estrogen nuclear receptor alpha ligand-binding 
                     domain"
                     /label=ER-LBD
                     /translation="KRSKKNSLALSLTADQMVSALLDAEPPILYSEYDPTRPFSEASMM
                     GLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEILMIGLVWRSMEHPGKL
                     LFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEFVCLKSIILLNSGVYTF
                     LSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRLAQLLLILSHIRHMSNK
                     GMEHLYSMKCKNVVPLYDLLLEMLDAHRLHAPTSRGGASVEETDQSHLATAGSTSSHSL
                     QKYYITGEAEGFPATV"
     polyA_signal    2838..2972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2875..2894)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2929..2948
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2982..3001)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2983..3001)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3088..3105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3259..3847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3339..3358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4018..4809,4810..4878))
                     /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     4641..4660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4879..4983)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5051..5069
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
       61 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
      121 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
      181 caccatatgg acatattgtc gttagaacgc ggctacaatt aatacataac cttatgtatc
      241 atacacatac gatttaggtg acactataga actcgatgtg gaatgtgtgt cagttagggt
      301 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
      361 cagcaaggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
      421 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
      481 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
      541 gcctcggcct ctgagctatt ccagaagtag tgaagaggct tttttggagg agatctaagc
      601 tagcgccgcc accatgggcc ctaaaaagaa gcgtaaagtc gcccccccga ccgatgtcag
      661 cctgggggac gagctccact tagacggcga ggacgtggcg atggcgcatg ccgacgcgct
      721 agacgatttc gatctggaca tgttggggga cggggattcc ccgggtccgg gatttacccc
      781 ccacgactcc gccccctacg gcgctctgga tatggccgac ttcgagtttg agcagatgtt
      841 taccgatgcc cttggaattg acgagtacgg tggggaattc atgaccatga ccctccacac
      901 caaagcatct gggatggccc tactgcatca gatccaaggg aacgagctgg agcccctgaa
      961 ccgtccgcag ctcaagatcc ccctggagcg gcccctgggc gaggtgtacc tggacagcag
     1021 caagcccgcc gtgtacaact accccgaggg cgccgcctac gagttcaacg ccgcggccgc
     1081 cgccaacgcg caggtctacg gtcagaccgg cctcccctac ggccccgggt ctgaggctgc
     1141 ggcgttcggc tccaacggcc tggggggttt ccccccactc aacagcgtgt ctccgagccc
     1201 gctgatgcta ctgcacccgc cgccgcagct gtcgcctttc ctgcagcccc acggccagca
     1261 ggtgccctac tacctggaga acgagcccag cggctacacg gtgcgcgagg ccggcccgcc
     1321 ggcattctac aggccaaatt cagataatcg acgccagggt ggcagagaaa gattggccag
     1381 taccaatgac aagggaagta tggctatgga atctgccaag gagactcgct actgtgcagt
     1441 gtgcaatgac tatgcttcag gctaccatta tggagtctgg tcctgtgagg gctgcaaggc
     1501 cttcttcaag agaagtattc aaggacataa cgactatatg tgtccagcca ccaaccagtg
     1561 caccattgat aaaaacagga ggaagagctg ccaggcctgc cggctccgca aatgctacga
     1621 agtgggaatg atgaaaggtg ggatacgaaa agaccgaaga ggagggagaa tgttgaaaca
     1681 caagcgccag agagatgatg gggagggcag gggtgaagtg gggtctgctg gagacatgag
     1741 agctgccaac ctttggccaa gcccgctcat gatcaaacgc tctaagaaga acagcctggc
     1801 cttgtccctg acggccgacc agatggtcag tgccttgttg gatgctgagc cccccatact
     1861 ctattccgag tatgatccta ccagaccctt cagtgaagct tcgatgatgg gcttactgac
     1921 caacctggca gacagggagc tggttcacat gatcaactgg gcgaagaggg tgccaggctt
     1981 tgtggatttg accctccatg atcaggtcca ccttctagaa tgtgcctggc tagagatcct
     2041 gatgattggt ctcgtctggc gctccatgga gcacccaggg aagctactgt ttgctcctaa
     2101 cttgctcttg gacaggaacc agggaaaatg tgtagagggc atggtggaga tcttcgacat
     2161 gctgctggct acatcatctc ggttccgcat gatgaatctg cagggagagg agtttgtgtg
     2221 cctcaaatct attattttgc ttaattctgg agtgtacaca tttctgtcca gcaccctgaa
     2281 gtctctggaa gagaaggacc atatccaccg agtcctggac aagatcacag acactttgat
     2341 ccacctgatg gccaaggcag gcctgaccct gcagcagcag caccagcggc tggcccagct
     2401 cctcctcatc ctctcccaca tcaggcacat gagtaacaaa ggcatggagc atctgtacag
     2461 catgaagtgc aagaacgtgg tgcccctcta tgacctgctg ctggagatgc tggacgccca
     2521 ccgcctacat gcgcccacta gccgtggagg ggcatccgtg gaggagacgg accaaagcca
     2581 cttggccact gcgggctcta cttcatcgca ttccttgcaa aagtattaca tcacggggga
     2641 ggcagagggt ttccctgcca cagtctgaga tcgaattcct gcagcccggg ggatccgtcg
     2701 acgcgtctgc agaagcttct agataagtaa tgatcataat cagccatatc acatctgtag
     2761 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     2821 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     2881 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     2941 aactcatcaa tgtatcttat catgtctgga tctgccggtc tccctatagt gagtcgtatt
     3001 aatttcgata agccaggtta acctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3061 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3121 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3181 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3241 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     3301 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     3361 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     3421 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     3481 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     3541 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     3601 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     3661 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     3721 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     3781 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     3841 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     3901 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     3961 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4021 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4081 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4141 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4201 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4261 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     4321 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     4381 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     4441 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     4501 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     4561 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     4621 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     4681 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     4741 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     4801 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     4861 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     4921 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     4981 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt
     5041 aacctataaa aataggcgta tcacgaggcc ctttcgt
//
