LOCUS       sequence-99647-e         942 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 942)
  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 942)
  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-8a
FEATURES             Location/Qualifiers
     source          1..942
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             25..47
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             66..83
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             93..113
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(126..128,129..842)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(168..189)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(429..448)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     776..797
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             895..912
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 taacaattcc cctctagaaa taattttgtt taactttaag aaggagatat catatgggca
       61 gcagccatca tcatcatcat cacagcagcg gcgaaaacct ttacttccag ggcgccatgg
      121 ccaaagtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      181 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      241 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      301 ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
      361 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      421 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
      481 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
      541 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
      601 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
      661 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
      721 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
      781 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
      841 aaggtaccgg atccgaattc gagctccgtc gacaagcttg cggccgcact cgagcaccac
      901 caccaccacc actgagatcc ggctgctaac aaagcccgaa ag
//
