LOCUS       sequence-99661-e        1066 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1066)
  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 1066)
  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-8b
FEATURES             Location/Qualifiers
     source          1..1066
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             2..24
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             43..60
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             70..90
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(106..108,109..822)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(148..169)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(409..428)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     756..777
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             875..892
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(945..963)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      959..1006
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttttgtttaa ctttaagaag gagatatacc atgggcagca gccatcatca tcatcatcac
       61 agcagcggcg aaaaccttta cttccagggc catatggcta gcaaagtgag caagggcgag
      121 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      181 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      241 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccttcggc
      301 tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag
      361 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      421 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      481 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      541 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      601 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      661 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
      721 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      781 gccgccggga tcactctcgg catggacgag ctgtacaagt aaggtaccgg atccgaattc
      841 gagctccgtc gacaagcttg cggccgcact cgagcaccac caccaccacc actgagatcc
      901 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
      961 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
     1021 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaa
//
