LOCUS       sequence-35219-e         865 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 865)
  TITLE     LIC N-terminal fusion vectors for E. coli expression
REFERENCE   2  (bases 1 to 865)
  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 ProteinG TEV LIC cloning vector (1P)
FEATURES             Location/Qualifiers
     source          1..865
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(21..38)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(104..124)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(134..295)
                     /codon_start=1
                     /product="B1 domain of Streptococcal protein G   "
                     /label=GB1
                     /note="effective as a solubilizing fusion partner (Cheng 
                     and Patel, 2004)"
                     /translation="YKLILNGKTLKGETTTEAVDAATAEKVFKQYANDNGVDGEWTYDD
                     ATKTFTVTE"
     CDS             complement(314..331)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             350..372
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    387..411
                     /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(411..430)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(412..430)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(514..533)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        739..816
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(836..855)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ttgttagcag ccggatctca gtggtggtgg tggtggtgct cgagtgcggc cgcaagcttg
       61 tcgacggcgc tcgaattcgg atccgttatc cacttccaat attggattgg aagtacaggt
      121 tttcctcgat cccttcagta actgtaaagg tcttagtcgc atcgtcgtaa gtccattcac
      181 cgtcaacacc gttgtcgtta gcgtattgtt tgaagacttt ttctgcagta gcagcatcaa
      241 cagcttcagt agttgtttcg cctttcaatg ttttaccatt aaggattaat ttgtaagtag
      301 tcatagaaga accatggtga tggtgatggt gagaagaacc catggtatat ctccttctta
      361 aagttaaaca aaattatttc tagaggggaa ttgttatccg ctcacaattc ccctatagtg
      421 agtcgtatta atttcgcggg atcgagatct cgatcctcta cgccggacgc atcgtggccg
      481 gcatcaccgg cgccacaggt gcggttgctg gcgcctatat cgccgacatc accgatgggg
      541 aagatcgggc tcgccacttc gggctcatga gcgcttgttt cggcgtgggt atggtggcag
      601 gccccgtggc cgggggactg ttgggcgcca tctccttgca tgcaccattc cttgcggcgg
      661 cggtgctcaa cggcctcaac ctactactgg gctgcttcct aatgcaggag tcgcataagg
      721 gagagcgtcg agatcccgga caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga
      781 tagcgcccgg aagagagtca attcagggtg gtgaatgtga aaccagtaac gttatacgat
      841 gtcgcagagt atgccggtgt ctctt
//
