LOCUS       sequence-99645-e        1608 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1608)
  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 1608)
  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-6a
FEATURES             Location/Qualifiers
     source          1..1608
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             1..21
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             35..52
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             71..721
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
     primer_bind     682..704
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             758..778
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(791..793,794..1507)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(833..854)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1094..1113)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1441..1462
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1560..1577
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ttgtttaact ttaagaagga gatataccat gaaacatcac catcaccatc acaacactag
       61 tagcaattcc atgtccccta tactaggtta ttggaaaatt aagggccttg tgcaacccac
      121 tcgacttctt ttggaatatc ttgaagaaaa atatgaagag catttgtatg agcgcgatga
      181 aggtgataaa tggcgaaaca aaaagtttga attgggtttg gagtttccca atcttcctta
      241 ttatattgat ggtgatgtta aattaacaca gtctatggcc atcatacgtt atatagctga
      301 caagcacaac atgttgggtg gttgtccaaa agagcgtgca gagatttcaa tgcttgaagg
      361 agcggttttg gatattagat acggtgtttc gagaattgca tatagtaaag actttgaaac
      421 tctcaaagtt gattttctta gcaagctacc tgaaatgctg aaaatgttcg aagatcgttt
      481 atgtcataaa acatatttaa atggtgatca tgtaacccat cctgacttca tgttgtatga
      541 cgctcttgat gttgttttat acatggaccc aatgtgcctg gatgcgttcc caaaattagt
      601 ttgttttaaa aaacgtattg aagctatccc acaaattgat aagtacttga aatccagcaa
      661 gtatatagca tggcctttgc agggctggca agccacgttt ggtggtggcg accatcctcc
      721 aactagtgga tctggtggtg gtggcggatc catgagcgag aatctttatt ttcagggcgc
      781 catggccaaa gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      841 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
      901 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
      961 gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca
     1021 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1081 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1141 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1201 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1261 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1321 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1381 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1441 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1501 caagtaaggt accggatccg aattcgagct ccgtcgacaa gcttgccgcc gcactcgagc
     1561 accaccacca ccaccactga gatccggctg ctaacaaagc ccgaaagg
//
