LOCUS       sequence-99652-e        1372 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1372)
  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 1372)
  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-14a
FEATURES             Location/Qualifiers
     source          1..1372
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             14..36
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             49..66
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             532..552
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(565..567,568..1281)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(607..628)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(868..887)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1215..1236
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1334..1351
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 cctctagaat aattttgttt aactttaaga aggagatata ctatgaaaca tcaccatcac
       61 catcacccta tgaccgacat ctacatcccg ccggagggtc tctacttccg cctccttggc
      121 tttgccagtc ggcaggtgat cttcgcgcgc aactctccct ctcccgatgt tggtctgtct
      181 ccggtcaacg accaggctac cgaccagtac ttctcgctca tctacggcac tggagaacac
      241 gccggtctct acgcgataaa gagcaaagcg acgggcaagg tgctcttctc gcgtaggcct
      301 gcggaaccgt atgtgggcca aatcgatggc gacgggcgtt atcccgacaa ctggttcaag
      361 attgagccag gaaagaccta tctctccaaa tatttccggc tcgttcagcc gtcgactggc
      421 accgcgcttg tctcgcgcac gcatttgcag ccatacttct ggaatcaccc tcagactgaa
      481 gtcttcgacg accaatactt caccttcctc ttcgaggatg caagctcttc tgaaaacctg
      541 tatttccagg gcgccatggc caaagtgagc aagggcgagg agctgttcac cggggtggtg
      601 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
      661 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
      721 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
      781 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
      841 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
      901 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
      961 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     1021 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     1081 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     1141 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     1201 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     1261 atggacgagc tgtacaagta aggtaccgga tccgaattcg agctccgtcg acaagcttgc
     1321 ggccgcactc gagcaccacc accaccacca ctgagatccg gctgctaaca aa
//
