LOCUS       sequence-50688-e         898 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 898)
  AUTHORS   Shen A, Lupardus PJ, Morell M, Ponder EL, Sadaghiani AM, Garcia KC, 
            Bogyo M
  TITLE     Simplified, enhanced protein purification using an inducible, 
            autoprocessing enzyme tag.
  JOURNAL   PLoS One. 2009 Dec 2;4(12):e8119.
  PUBMED    19956581
REFERENCE   2  (bases 1 to 898)
  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: "PLoS One.";
            date: "2009-12-2"; volume: "4(12)"; pages: "e8119"
COMMENT     Sequence Label: pET28a-CPDSalI
FEATURES             Location/Qualifiers
     source          1..898
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(17..34)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(702..734)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(738..755)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(765..782)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             801..823
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    838..862
                     /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(862..881)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(863..881)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tagcagccgg atctcagtgg tggtggtggt ggtgctcgag accttgcgcg tcccagctta
       61 gcgaaacttt gttgttttct gccttttgaa cccaatcgcc attcgcgtcc ttggtatgct
      121 tacgtcccgc ctcgtctacg gccagttcag aactacgaac agagacatcg acacgaagac
      181 cattcgcatc catcgcgtta ataaactgat gaccaaagcc tttttgcttg tcgtcactca
      241 ccaaagaaca accaacaata ctgatgtgat ccggtttgtt gttgatgttt tcggcttgat
      301 taaacgactg ttggaacttg gccaatttca cggccaactc atcggcactg taaccactta
      361 agcgagtatt gttagtttct gagtggtcgc gaccatgccc caccaactgc caacgtagct
      421 ttccatccag ttttgacgga tcgccataca ccacgcgata gttgccgtct gaatcgagct
      481 gcaccaccac actgctttca gcatgtttac ctgctaaatt ggctgccgct tttgctacta
      541 ccgggtcgtt ttccatttga acgatgattt gaccgtcgaa gcgggtttca ccaccatctg
      601 tcgttggtgt aaccgtaatc gggccccagc tattaacatt ttgattatgg agtatttttc
      661 catccgctaa tgcgtcgacg gagctcgaat tcggatccgc gacccatttg ctgtccacca
      721 gtcatgctag ccatatggct gccgcgcggc accaggccgc tgctgtgatg atgatgatga
      781 tggctgctgc ccatggtata tctccttctt aaagttaaac aaaattattt ctagagggga
      841 attgttatcc gctcacaatt cccctatagt gagtcgtatt aatttcgcgg gatcgaga
//
