LOCUS       sequence-150165-         862 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression plasmid used for T7 promoter driven expression 
            of recombinant proteins with N-terminal 8X His, thrombin cleavage 
            site, SUMO, and TEV cleavage site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 862)
  TITLE     SGC Toronto Q3 Q4 2015, Q1 2016
REFERENCE   2  (bases 1 to 862)
  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: pET28-MKH8SUMO
FEATURES             Location/Qualifiers
     source          1..862
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             31..54
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             64..81
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             82..375
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             379..399
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    complement(427..460)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 actttaagaa ggagatatac catgaaaaaa catcaccatc atcatcatca tcacagcagc
       61 ggcctggttc cgcgtggtag tatgtcggac tcagaagtca atcaagaagc taagccagag
      121 gtcaagccag aagtcaagcc tgagactcac atcaatttaa aggtgtccga tggatcttca
      181 gagatcttct tcaagatcaa aaagaccact cctttaagaa ggctgatgga agcgttcgct
      241 aaaagacagg gtaaggaaat ggactcctta agattcttgt acgacggtat tagaatccaa
      301 gctgatcaga cccctgaaga tttggacatg gaggataacg atattattga ggctcacaga
      361 gaacagattg gtggaacaga aaacttgtat ttccagggcg gagaccgttc tcatcctgaa
      421 agatccataa cttcgtatag catacattat acgaagttat gcggccgcga cgtccacata
      481 tacctgccgt tcactattat ttagtgaaat gagatattat gatattttct gaattgtgat
      541 taaaaaggca actttatgcc catgcaacag aaactataaa aaatacagag aatgaaaaga
      601 aacagataga ttttttagtt ctttaggccc gtagtctgca aatcctttta tgattttcta
      661 tcaaacaaaa gaggaaaata gaccagttgc aatccaaacg agagtctaat agaatgaggt
      721 cgaaaagtaa atcgcgcggg tttgttactg ataaagcagg caagacctaa aatgtgtaaa
      781 gggcaaagtg tatactttgg cgtcacccct tacatatttt aggtcttttt ttattgtgcg
      841 taactaactt gccatcttca aa
//
