LOCUS       sequence-70898-e         938 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  use to create N-terminal PelB-Twin-Strep-tag fusion proteins or dual
            tagged fusion with N-terminal PelB-Twin-Strep-tag and C-terminal 6x 
            His-tag for periplasmic translocation via PelB signal sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 938)
  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 938)
  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: pTDpelB-NTwinStrep
FEATURES             Location/Qualifiers
     source          1..938
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        join(342..347,348..364,365..371)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    379..395
                     /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)."
     sig_peptide     443..508
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             join(509..514,515..538,539..574,575..598)
                     /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             695..712
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      764..858
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 aaagaaaaac caccctggcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
       61 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
      121 attaatgtaa gttagcgcga attgatctgg tttgacagct tatcatcgac tgcacggtgc
      181 accaatgctt ctggcgtcag gcagccatcg gaagctgtgg tatggctgtg caggtcgtaa
      241 atcactgcat aattcgtgtc gctcaaggcg cactcccgtt ctggataatg ttttttgcgc
      301 cgacatcata acggttctgg caaatattct gaaatgagct gttgacaatt aatcatccgg
      361 ctcgtataat gtgtggaatt gtgagcggat aacaatttca caccctaggc cgcggccgcg
      421 cgaattggaa ggagatatac aaatgaagta cctgctgccg accgccgccg ccggcctgct
      481 gctgctggcc gcccagccgg ccatggccag cgcttggagc cacccgcagt tcgagaaagg
      541 tggaggttcc ggaggtggat cgggaggttc ggcgtggagc cacccgcagt tcgaaaaagg
      601 cgccgagacc gcggtcccga attcgagctc ggtacccggg gatccctcga ggtcgacctg
      661 cagggggacc atggtctcag gcctgagagg atcgcatcac catcaccatc actaaaagct
      721 tgcggccgcg tcgtgactgg gaaaaccctg gcgactagtc ttggactcct gttgatagat
      781 ccagtaatga cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg
      841 ttttttattg gtgagaatcc aggggtcccc aataattacg atttacgtat ttaaatgaac
      901 cttgaccgaa cgcagcggtg gtaacggcgc agtggcgg
//
