LOCUS       sequence-35235-e         825 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 825)
  TITLE     2-series LIC N-terminal fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 825)
  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"
COMMENT     Sequence Label: pET Flag TEV LIC cloning vector (2L-T)
FEATURES             Location/Qualifiers
     source          1..825
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     55..74
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        55..73
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             162..184
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             195..218
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             219..239
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     primer_bind     complement(364..382)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      378..425
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(791..796,797..813,814..819))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
ORIGIN
        1 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
       61 gactcactat agggagacca caacggtttc cctctagtgc cggctccgga gagctcttta
      121 attaagcggc cgccctgcag gactcgagtt ctagaaataa ttttgtttaa ctttaagaag
      181 gagatataca tatggactac aaagacgatg acgacaagga aaacctgtac ttccaatcca
      241 atattggaag tggataacgg atccgcgatc gcggcgcgcc acctggtggc cggccggtac
      301 cacgcgtgcg cgctgatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
      361 ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt
      421 ttttgctgaa aggaggaact atatccggat atccacagga cgggtgtggt cgccatgatc
      481 gcgtagtcga tagtggctcc aagtagcgaa gcgagcagga ctgggcggcg gccaaagcgg
      541 tcggacagtg ctccgagaac gggtgcgcat agaaattgca tcaacgcata tagcgctagc
      601 agcacgccat agtgactggc gatgctgtcg gaatggacga tatcccgcaa gaggcccggc
      661 agtaccggca taaccaagcc tatgcctaca gcatccaggg tgacggtgcc gaggatgacg
      721 atgagcgcat tgttagattt catacacggt gcctgactgc gttagcaatt taactgtgat
      781 aaactaccgc attaaagctt atcgatgata agctgtcaaa catga
//
