LOCUS       sequence-103889-         685 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  E. coli expression vector with N-terminal Trx-tag, 6X His-tag, 
            thrombin site, S-tag, and TEV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 685)
  TITLE     Arrowsmith Lab Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 685)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET32a-LIC
FEATURES             Location/Qualifiers
     source          1..685
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             11..337
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             359..376
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             386..403
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             410..454
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             470..490
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     protein_bind    complement(518..551)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 agatatacat atgagcgata aaattattca cctgactgac gacagttttg acacggatgt
       61 actcaaagcg gacggggcga tcctcgtcga tttctgggca gagtggtgcg gtccgtgcaa
      121 aatgatcgcc ccgattctgg atgaaatcgc tgacgaatat cagggcaaac tgaccgttgc
      181 aaaactgaac atcgatcaaa accctggcac tgcgccgaaa tatggcatcc gtggtatccc
      241 gactctgctg ctgttcaaaa acggtgaagt ggcggcaacc aaagtgggtg cactgtctaa
      301 aggtcagttg aaagagttcc tcgacgctaa cctggccggt tctggttctg gccatatgca
      361 ccatcatcat catcattctt ctggtctggt gccacgcggt tctggtatga aagaaaccgc
      421 tgctgctaaa ttcgaacgcc agcacatgga cagcccagat ctgggtaccg aaaacttgta
      481 tttccagagc catatgagtt ctcctcctga aagatccata acttcgtata gcatacatta
      541 tacgaagtta tgcggccgcg acgtccacat atacctgccg ttcactatta tttagtgaaa
      601 tgagatatta tgatattttc tgaattgtga ttaaaaaggc aactttatgc ccatgcaaca
      661 gaaactataa aaaatacaga gaatg
//
