LOCUS       sequence-99659-e        1612 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1612)
  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 1612)
  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-6b
FEATURES             Location/Qualifiers
     source          1..1612
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             34..51
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             70..720
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPP"
     primer_bind     681..703
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             757..777
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(793..795,796..1509)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(835..856)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1096..1115)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1443..1464
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1562..1579
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tgtttaactt taagaaggag atataccatg aaacatcacc atcaccatca caacactagt
       61 agcaattcca tgtcccctat actaggttat tggaaaatta agggccttgt gcaacccact
      121 cgacttcttt tggaatatct tgaagaaaaa tatgaagagc atttgtatga gcgcgatgaa
      181 ggtgataaat ggcgaaacaa aaagtttgaa ttgggtttgg agtttcccaa tcttccttat
      241 tatattgatg gtgatgttaa attaacacag tctatggcca tcatacgtta tatagctgac
      301 aagcacaaca tgttgggtgg ttgtccaaaa gagcgtgcag agatttcaat gcttgaagga
      361 gcggttttgg atattagata cggtgtttcg agaattgcat atagtaaaga ctttgaaact
      421 ctcaaagttg attttcttag caagctacct gaaatgctga aaatgttcga agatcgttta
      481 tgtcataaaa catatttaaa tggtgatcat gtaacccatc ctgacttcat gttgtatgac
      541 gctcttgatg ttgttttata catggaccca atgtgcctgg atgcgttccc aaaattagtt
      601 tgttttaaaa aacgtattga agctatccca caaattgata agtacttgaa atccagcaag
      661 tatatagcat ggcctttgca gggctggcaa gccacgtttg gtggtggcga ccatcctcca
      721 actagtggat ctggtggtgg tggcggatcc atgagcgaga atctttattt tcagggccat
      781 atggctagca aagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      841 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
      901 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      961 tggcccaccc tcgtgaccac cttcggctac ggcctgcagt gcttcgcccg ctaccccgac
     1021 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     1081 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     1141 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     1201 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     1261 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     1321 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     1381 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     1441 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     1501 tacaagtaag gtaccggatc cgaattcgag ctccgtcgac aagcttgcgg ccgcactcga
     1561 gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg ag
//
