LOCUS       sequence-99666-e        1385 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1385)
  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 1385)
  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-14b
FEATURES             Location/Qualifiers
     source          1..1385
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             28..50
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             63..80
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             546..566
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(582..584,585..1298)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(624..645)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(885..904)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1232..1253
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1351..1368
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 agcgtaaaca ttccctctag aataattttt gtttaacttt aagaaggaga tatactatga
       61 aacatcacca tcaccatcac cctatgaccg acatctacat cccgccggag ggtctctact
      121 tccgcctcct tggctttgcc agtcggcagg tgatcttcgc gcgcaactct ccctctcccg
      181 atgttggtct gtctccggtc aacgaccagg ctaccgacca gtacttctcg ctcatctacg
      241 gcactggaga acacgccggt ctctacgcga taaagagcaa agcgacgggc aaggtgctct
      301 tctcgcgtag gcctgcggaa ccgtatgtgg gccaaatcga tggcgacggg cgttatcccg
      361 acaactggtt caagattgag ccaggaaaga cctatctctc caaatatttc cggctcgttc
      421 agccgtcgac tggcaccgcg cttgtctcgc gcacgcattt gcagccatac ttctggaatc
      481 accctcagac tgaagtcttc gacgaccaat acttcacctt cctcttcgag gatgcaagct
      541 cttctgaaaa cctgtatttc cagggccata tggctagcaa agtgagcaag ggcgaggagc
      601 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      661 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      721 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg
      781 gcctgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      841 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      901 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      961 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     1021 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     1081 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1141 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     1201 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1261 ccgggatcac tctcggcatg gacgagctgt acaagtaagg taccggatcc gaattcgagc
     1321 tccgtcgaca agcttgcggc cgcactcgag caccaccacc accaccactg agatccggct
     1381 gctaa
//
