LOCUS       sequence-35221-e         889 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 889)
  TITLE     LIC N-terminal fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 889)
  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 (1L)
FEATURES             Location/Qualifiers
     source          1..889
                     /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(129..152)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     RBS             162..184
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    199..223
                     /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(223..242)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(224..242)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(326..345)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        551..628
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(648..667)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggctttgtta gcagccggat ctcagtggtg gtggtggtgg tgctcgagtg cggccgcaag
       61 cttgtcgacg gcgctcgaat tcggatccgt tatccacttc caatattgga ttggaagtac
      121 aggttttcct tgtcgtcatc gtctttgtag tccatggtat atctccttct taaagttaaa
      181 caaaattatt tctagagggg aattgttatc cgctcacaat tcccctatag tgagtcgtat
      241 taatttcgcg ggatcgagat ctcgatcctc tacgccggac gcatcgtggc cggcatcacc
      301 ggcgccacag gtgcggttgc tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg
      361 gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg gtatggtggc aggccccgtg
      421 gccgggggac tgttgggcgc catctccttg catgcaccat tccttgcggc ggcggtgctc
      481 aacggcctca acctactact gggctgcttc ctaatgcagg agtcgcataa gggagagcgt
      541 cgagatcccg gacaccatcg aatggcgcaa aacctttcgc ggtatggcat gatagcgccc
      601 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
      661 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
      721 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
      781 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
      841 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgcc
//
