LOCUS       sequence-101338-         790 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 790)
  AUTHORS   Goulas T, Cuppari A, Garcia-Castellanos R, Snipas S, Glockshuber R, 
            Arolas JL, Gomis-Ruth FX
  TITLE     The pCri System: A Vector Collection for Recombinant Protein 
            Expression and Purification.
  JOURNAL   PLoS One. 2014 Nov 11;9(11):e112643. doi: 
            10.1371/journal.pone.0112643. eCollection 2014.
  PUBMED    25386923
REFERENCE   2  (bases 1 to 790)
  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: "PLoS One.";
            date: "2014-11-11"; volume: "9(11):e112643. doi"; pages: " 
            10.1371/journal.pone.0112643. eCollection 2014"
COMMENT     Sequence Label: pCri-4b
FEATURES             Location/Qualifiers
     source          1..790
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     184..203
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     287..306
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        287..305
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    306..330
                     /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)."
     RBS             345..367
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             380..397
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             401..727
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             746..766
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
ORIGIN
        1 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
       61 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
      121 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
      181 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
      241 cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact
      301 ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag
      361 aaggagatat accatgaaac atcaccatca ccatcacccc atgagcgata aaattattca
      421 cctgactgac gacagttttg acacggatgt actcaaagcg gacggggcga tcctcgtcga
      481 tttctgggca gagtggtgcg gtccgtgcaa aatgatcgcc ccgattctgg atgaaatcgc
      541 tgacgaatat cagggcaaac tgaccgttgc aaaactgaac atcgatcaaa accctggcac
      601 tgcgccgaaa tatggcatcc gtggtatccc gactctgctg ctgttcaaaa acggtgaagt
      661 ggcggcaacc aaagtgggtg cactgtctaa aggtcagttg aaagagttcc tcgacgctaa
      721 cctggccgga tctggcagtg gttctgagaa tctttatttt cagggcgccc atatggctag
      781 caaagtgagc
//
