LOCUS       sequence-70757-e         960 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  use to create N-terminal PelB and C-terminal Twin-Strep-tagged 
            fusion proteins for periplasmic translocation via PelB signal 
            sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 960)
  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 960)
  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-CTwinStrep
FEATURES             Location/Qualifiers
     source          1..960
                     /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     453..518
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             join(564..569,570..593,594..629,630..653)
                     /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"
     terminator      711..805
                     /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 cgaattgtta actttaagaa ggagatatac atatgaagta cctgctgccg accgccgccg
      481 ccggcctgct gctgctggcc gcccagccgg ccatggccga tccgaattcg agctccgtcg
      541 actctagagc ggccgcactc gagagcgctt ggagccaccc gcagttcgag aaaggtggag
      601 gttccggagg tggatcggga ggttcggcgt ggagccaccc gcagttcgaa aaataatgat
      661 gaaagcttgc ggccgcgtcg tgactgggaa aaccctggcg actagtcttg gactcctgtt
      721 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
      781 ccgggcgttt tttattggtg agaatccagg ggtccccaat aattacgatt tacgtattta
      841 aatgaacctt gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt
      901 atgactgttt ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
//
