LOCUS       sequence-97671-e         915 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pET21-DTag encodes Dead streptavidin (negligible biotin binding) 
            bearing a C-terminal SpyTag, for generation of chimeric SpyAvidin 
            tetramers.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 915)
  AUTHORS   Fairhead M, Veggiani G, Lever M, Yan J, Mesner D, Robinson CV, 
            Dushek O, van der Merwe PA, Howarth M
  TITLE     SpyAvidin hubs enable precise and ultrastable orthogonal 
            nanoassembly.
  JOURNAL   J Am Chem Soc. 2014 Sep 3;136(35):12355-63. doi: 10.1021/ja505584f. 
            Epub 2014 Aug 21.
  PUBMED    25111182
REFERENCE   2  (bases 1 to 915)
  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: "10.1021/ja505584f";
            journalName: "J Am Chem Soc"; date: "2014-09-3- 3"; volume: "136"; 
            issue: "35"; pages: "12355-63"
COMMENT     Sequence Label: Dead Streptavidin-SpyTag
FEATURES             Location/Qualifiers
     source          1..915
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(15..32)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             505..527
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    542..566
                     /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(566..585)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(567..585)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(669..688)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcagccggat ctcagtggtg gtggtggtgg tgctcgagtg cggccgcaag cttttattac
       61 gtaggtttgt aggcatccac catcacgatg tgggcgccgc ctgatcctga tccggaagca
      121 gcggacggtt taactttggt gaaggtgtcg tgaccaacca gggtggattt ccacgcgtta
      181 gcttcggtgg tgccggaggt caacaaccac tgggtgttga tacgagcttc agcaccacca
      241 acgtactggc cagaccaggt ggtagcggag tgagcgttac ggtagttgtt tttccaagca
      301 acggtccaac ccagagcggt tccggaaccg tcggtagccg gagcggagtc gtaacgaccg
      361 gtcagaacgt atctagattc agcgttacca acagcggctt cgtaggtacc ggtcagagca
      421 ccgtcagcac cagcggtaac gatgaaggtg tctcccagct gggcgtacca ggtgccggtg
      481 ataccagctt cagccatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag
      541 gggaattgtt atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga
      601 gatctcgatc ctctacgccg gacgcatcgt ggccggcatc accggcgcca caggtgcggt
      661 tgctggcgcc tatatcgccg acatcaccga tggggaagat cgggctcgcc acttcgggct
      721 catgagcgct tgtttcggcg tgggtatggt ggcaggcccc gtggccgggg gactgttggg
      781 cgccatctcc ttgcatgcac cattccttgc ggcggcggtg ctcaacggcc tcaacctact
      841 actgggctgc ttcctaatgc aggagtcgca taagggagag cgtcgagatc ccggacacca
      901 tcgaatggcg caaaa
//
