LOCUS       sequence-135965-         878 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  pET28a modified to contain TEV protease fusion tag instead of T7 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 878)
  AUTHORS   Kappock
  TITLE     Vectors
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 878)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pJK635
FEATURES             Location/Qualifiers
     source          1..878
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(26..43)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(93..125)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(129..149)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             complement(174..191)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             204..226
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    241..265
                     /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(265..284)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(266..284)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(368..387)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        593..670
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(690..709)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 gggctttgtt agcagccgga tctcagtggt ggtggtggtg gtgctcgagt gcggccgcaa
       61 gcttgtcgac ggagctcgaa ttcggatccg cgacccattt gctgtccacc agtcatgcta
      121 gccatatgag actgaaagta cagattttcg gtgcccagat ccacgccgct tgaatgatgg
      181 tgatgatggt gtgccatggt atatctcctt cttaaagtta aacaaaatta tttctagagg
      241 ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg cgggatcgag
      301 atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac aggtgcggtt
      361 gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca cttcgggctc
      421 atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg actgttgggc
      481 gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct caacctacta
      541 ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc cggacaccat
      601 cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga gtcaattcag
      661 ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg gtgtctctta
      721 tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa
      781 agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac aacaactggc
      841 gggcaaacag tcgttgctga ttggcgttgc cacctcca
//
