LOCUS       sequence-99669-e        1281 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1281)
  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 1281)
  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-4a-Strep
FEATURES             Location/Qualifiers
     source          1..1281
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             36..53
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             57..383
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             402..422
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(435..437,438..1151)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(477..498)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(738..757)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1085..1106
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1204..1227
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1237..1254
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tttgtttaac tttaagaagg agatatacca tgaaacatca ccatcaccat caccccatga
       61 gcgataaaat tattcacctg actgacgaca gttttgacac ggatgtactc aaagcggacg
      121 gggcgatcct cgtcgatttc tgggcagagt ggtgcggtcc gtgcaaaatg atcgccccga
      181 ttctggatga aatcgctgac gaatatcagg gcaaactgac cgttgcaaaa ctgaacatcg
      241 atcaaaaccc tggcactgcg ccgaaatatg gcatccgtgg tatcccgact ctgctgctgt
      301 tcaaaaacgg tgaagtggcg gcaaccaaag tgggtgcact gtctaaaggt cagttgaaag
      361 agttcctcga cgctaacctg gccggatctg gcagtggttc tgagaatctt tattttcagg
      421 gcgccatggc caaagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      481 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      541 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      601 cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg
      661 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
      721 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
      781 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
      841 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
      901 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
      961 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1021 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1081 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1141 tgtacaagta aggtaccgga tccgaattcg agctccgtcg acaagcttgc ggccgcactc
     1201 gagtggagcc atccacaatt cgaaaagtaa actagtcacc accaccacca ccactgagat
     1261 ccggctgcta acaaagcccg a
//
