LOCUS       sequence-103166-         858 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of human p53 (1-73) (P27A) in e. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 858)
  AUTHORS   Borcherds W, Theillet FX, Katzer A, Finzel A, Mishall KM, Powell AT,
            Wu H, Manieri W, Dieterich C, Selenko P, Loewer A, Daughdrill GW
  TITLE     Disorder and residual helicity alter p53-Mdm2 binding affinity and 
            signaling in cells.
  JOURNAL   Nat Chem Biol. 2014 Dec;10(12):1000-2. doi: 10.1038/nchembio.1668. 
            Epub 2014 Nov 2.
  PUBMED    25362358
REFERENCE   2  (bases 1 to 858)
  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/nchembio.1668"; journalName: "Nat Chem Biol"; date: 
            "2014-12"; volume: "10"; issue: "12"; pages: "1000-2"
COMMENT     Sequence Label: pET28A-p53 (1-73) (P27A)
FEATURES             Location/Qualifiers
     source          1..858
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(18..35)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(271..288)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(298..315)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             334..356
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    371..395
                     /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)."
     primer_bind     complement(395..414)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(396..414)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(498..517)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        723..800
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(820..839)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttagcagccg gatctcagtg gtggtggtgg tggtgctcga gtcacacgcg gggtagcagc
       61 ctctggcatt ctgggagctt catctggacc tgggtcttca gtgaaccatt gttcaatatc
      121 gtccggggac agcatcaaat catccattgc ttgggacggc aagggggaca gaacgttgtt
      181 ttcagcaagt agtttccata ggtctgaaaa tgtttcctga ctcagagggg gctcgacgct
      241 aggatctgac tgcggctcct ccatcatatg gctgccgcgc ggcaccaggc cgctgctgtg
      301 atgatgatga tgatggctgc tgcccatggt atatctcctt cttaaagtta aacaaaatta
      361 tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg
      421 cgggatcgag atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
      481 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
      541 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
      601 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
      661 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc
      721 cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
      781 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
      841 gtgtctctta tcagaccg
//
