LOCUS       sequence-103487-         732 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of human p53 (1-73) (K24N) in e. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 732)
  AUTHORS   Zhan YA, Wu H, Powell AT, Daughdrill GW, Ytreberg FM
  TITLE     Impact of the K24N mutation on the transactivation domain of p53 and
            its binding to murine double-minute clone 2.
  JOURNAL   Proteins. 2013 Oct;81(10):1738-47. doi: 10.1002/prot.24310. Epub 
            2013 Jul 22.
  PUBMED    23609977
REFERENCE   2  (bases 1 to 732)
  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: 
            "10.1002/prot.24310"; journalName: "Proteins"; date: "2013-10"; 
            volume: "81"; issue: "10"; pages: "1738-47"
COMMENT     Sequence Label: pET28-p53 (1-73) (K24N)
FEATURES             Location/Qualifiers
     source          1..732
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(66..113)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     109..127
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(180..197)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(432..449)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(459..476)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             495..517
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    532..556
                     /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(556..575)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(557..575)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(659..678)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcgcttaatg cgccgctaca gggcgcgctc ccattcgcca atccggatat agttcctcct
       61 ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc
      121 tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt tgttagcagc cggatctcag
      181 tggtggtggt ggtggtgctc gagtcacacg cggggagcag cctctggcat tctgggagct
      241 tcatctggac ctgggtcttc agtgaaccat tgttcaatat cgtccgggga cagcatcaaa
      301 tcatccattg cttgggacgg caagggggac agaacgttgt tttcaggaag tagattccat
      361 aggtctgaaa atgtttcctg actcagaggg ggctcgacgc taggatctga ctgcggctcc
      421 tccatcatat ggctgccgcg cggcaccagg ccgctgctgt gatgatgatg atgatggctg
      481 ctgcccatgg tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt
      541 atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga gatctcgatc
      601 ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt tgctggcgcc
      661 tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct catgagcgct
      721 tgtttcggcg tg
//
