LOCUS       sequence-70752-e         976 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  use to create N-terminal Twin-Strep-tag fusion proteins or dual 
            tagged fusion with N-terminal Twin-Strep-tag and C-terminal 6x 
            His-tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 976)
  AUTHORS   Dammeyer T, Timmis KN, Tinnefeld P
  TITLE     Broad host range vectors for expression of proteins with (Twin-) 
            Strep-tag, His-tag and engineered, export optimized yellow 
            fluorescent protein.
  JOURNAL   Microb Cell Fact. 2013 May 20;12(1):49.
  PUBMED    23687945
REFERENCE   2  (bases 1 to 976)
  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: "Microb Cell
            Fact."; date: "2013-05-20"; volume: "12(1)"; pages: "49"
COMMENT     Sequence Label: pTD-NTwinStrep_Km
FEATURES             Location/Qualifiers
     source          1..976
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        join(357..362,363..379,380..386)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    394..410
                     /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)."
     CDS             join(464..469,470..493,494..529,530..553)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             650..667
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      719..813
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(934..953)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ccgtctcact ggtgaaaaga aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc
       61 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
      121 agtgagcgca acgcaattaa tgtaagttag cgcgaattga tctggtttga cagcttatca
      181 tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc tgtggtatgg
      241 ctgtgcaggt cgtaaatcac tgcataattc gtgtcgctca aggcgcactc ccgttctgga
      301 taatgttttt tgcgccgaca tcataacggt tctggcaaat attctgaaat gagctgttga
      361 caattaatca tccggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacaccc
      421 taggccgcgg ccgcgcgaat tggaaggaga tatacaaatg gctagcgctt ggagccaccc
      481 gcagttcgag aaaggtggag gttccggagg tggatcggga ggttcggcgt ggagccaccc
      541 gcagttcgaa aaaggcgccg agaccgcggt cccgaattcg agctcggtac ccggggatcc
      601 ctcgaggtcg acctgcaggg ggaccatggt ctcaggcctg agaggatcgc atcaccatca
      661 ccatcactaa aagcttgcgg ccgcgtcgtg actgggaaaa ccctggcgac tagtcttgga
      721 ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt cagaacgctc
      781 ggttgccgcc gggcgttttt tattggtgag aatccagggg tccccaataa ttacgattta
      841 aatttgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa
      901 caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc catattcagc
      961 gtgaaacgag ctgtag
//
