LOCUS       sequence-99658-e        1246 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Protein expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1246)
  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 1246)
  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-4b
FEATURES             Location/Qualifiers
     source          1..1246
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             32..49
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             53..379
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             398..418
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             join(437..439,440..1153)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(479..500)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(740..759)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1087..1108
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1206..1223
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 tttaacttta agaaggagat ataccatgaa acatcaccat caccatcacc ccatgagcga
       61 taaaattatt cacctgactg acgacagttt tgacacggat gtactcaaag cggacggggc
      121 gatcctcgtc gatttctggg cagagtggtg cggtccgtgc aaaatgatcg ccccgattct
      181 ggatgaaatc gctgacgaat atcagggcaa actgaccgtt gcaaaactga acatcgatca
      241 aaaccctggc actgcgccga aatatggcat ccgtggtatc ccgactctgc tgctgttcaa
      301 aaacggtgaa gtggcggcaa ccaaagtggg tgcactgtct aaaggtcagt tgaaagagtt
      361 cctcgacgct aacctggccg gatctggcag tggttctgag aatctttatt ttcagggcgc
      421 ccatatggct agcaaagtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      481 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      541 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      601 gccctggccc accctcgtga ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc
      661 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      721 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      781 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      841 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
      901 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
      961 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1021 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1081 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1141 gctgtacaag taaggtaccg gatccgaatt cgagctccgt cgacaagaaa gcggccgcac
     1201 tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagc
//
