LOCUS       sequence-35218-e         849 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 849)
  TITLE     LIC N-terminal fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 849)
  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 His6 Sumo TEV LIC cloning vector (1S)
FEATURES             Location/Qualifiers
     source          1..849
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(25..42)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(108..128)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(135..428)
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             complement(447..464)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             483..505
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    520..544
                     /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)."
     primer_bind     complement(544..563)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(545..563)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(647..666)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 ggctttgtta gcagccggat ctcagtggtg gtggtggtgg tgctcgagtg cggccgcaag
       61 cttgtcgacg gcgctcgaat tcggatccgt tatccacttc caatattgga ttggaagtac
      121 aggttttcct cgatcccacc aatctgttct ctgtgagcct caataatatc gttatcctcc
      181 atgtccaaat cttcaggggt ctgatcagct tgaattctaa taccgtcgta caagaatctt
      241 aaggagtcca tttccttacc ctgtctttta gcgaacgctt ccatcagcct tcttaaagga
      301 gtggtctttt tgatcttgaa gaagatctct gaagatccat cggacacctt taaattgatg
      361 tgagtctcag gcttgacttc tggcttgacc tctggcttag cttcttgatt gacttctgag
      421 tccgacatgc tagccataga agaaccatgg tgatggtgat ggtgagaaga acccatggta
      481 tatctccttc ttaaagttaa acaaaattat ttctagaggg gaattgttat ccgctcacaa
      541 ttcccctata gtgagtcgta ttaatttcgc gggatcgaga tctcgatcct ctacgccgga
      601 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
      661 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
      721 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctcctt gcatgcacca
      781 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
      841 gagtcgcat
//
