LOCUS       sequence-99644-e        1243 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1243)
  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 1243)
  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
FEATURES             Location/Qualifiers
     source          1..1243
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             26..43
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             47..373
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             392..412
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(425..427,428..1141)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(467..488)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(728..747)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1075..1096
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1194..1211
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tttaagaagg agatatacca tgaaacatca ccatcaccat caccccatga gcgataaaat
       61 tattcacctg actgacgaca gttttgacac ggatgtactc aaagcggacg gggcgatcct
      121 cgtcgatttc tgggcagagt ggtgcggtcc gtgcaaaatg atcgccccga ttctggatga
      181 aatcgctgac gaatatcagg gcaaactgac cgttgcaaaa ctgaacatcg atcaaaaccc
      241 tggcactgcg ccgaaatatg gcatccgtgg tatcccgact ctgctgctgt tcaaaaacgg
      301 tgaagtggcg gcaaccaaag tgggtgcact gtctaaaggt cagttgaaag agttcctcga
      361 cgctaacctg gccggatctg gcagtggttc tgagaatctt tattttcagg gcgccatggc
      421 caaagtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      481 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      541 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      601 cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg accacatgaa
      661 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      721 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      781 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      841 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
      901 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
      961 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1021 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1081 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1141 aggtaccgga tccgaattcg agctccgtcg acaagcttgc ggccgcactc gagcaccacc
     1201 accaccacca ctgagatccg gctgctaaca aagcccgaaa gaa
//
