LOCUS       sequence-21577-e         636 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  tandem-affinity purification plasmid; streptavidin-(streptag) and 
            calmodulin-(CBP) binding affinity tags.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 636)
  AUTHORS   Angers S, Thorpe CJ, Biechele TL, Goldenberg SJ, Zheng N, MacCoss 
            MJ, Moon RT
  TITLE     The KLHL12-Cullin-3 ubiquitin ligase negatively regulates the 
            Wnt-beta-catenin pathway by targeting Dishevelled for degradation.
  JOURNAL   Nat Cell Biol. 2006 Apr . 8(4):348-57.
  PUBMED    16547521
REFERENCE   2  (bases 1 to 636)
  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: "Nat Cell 
            Biol. 2006 Apr . 8(4):348-57."
COMMENT     Sequence Label: V51 pIRESpuro-GLUE (pGLUE)
FEATURES             Location/Qualifiers
     source          1..636
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     52..71
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        52..70
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      112..121
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             118..231
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             250..270
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             277..303
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             277..303
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             304..381
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
ORIGIN
        1 agcagagctc tctggctact agagaacccc tgcttactgg cttatcgaaa ttaatacgac
       61 tcactatagg gagacccaag cttggtaccg agctcggatc gatttaatta agccaccatg
      121 gacgagaaga ccaccggctg gcggggcggc cacgtggtgg agggcctggc cggcgagctg
      181 gagcagctgc gggcccggct ggagcaccac ccccagggcc agcgggagcc cggcggaagc
      241 cccggtggcg agaacctgta cttccagggc gcaagctacc cctacgacgt gcccgactac
      301 gccaagcggc ggtggaagaa gaacttcatc gccgtgagcg ccgccaaccg gttcaagaag
      361 atcagcagca gcggcgccct gattgatggg ggatctggcc ccggcggcgg cgcgcctgta
      421 cagatatctg cggccgcgtc gacggaattc gcccttttcg gatccactag taacggccgc
      481 cagtgtgctg gaattaattc gctgtctgcg agggccagct gttggggtga gtactccctc
      541 tcaaaagcgg gcatgacttc tgcgctaana ttgtcagttt ccaaaaacga ggaggatttg
      601 atattcacct ggcccgcggt gatgcctttg agggtg
//
