LOCUS       sequence-309687-         226 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For the protein expression with ALFA tag at C-terminus in mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 226)
  AUTHORS   Jin F, Shen C, Wang Y, Wang M, Sun M, Hattori M
  TITLE     Fluorescence-detection size-exclusion chromatography utilizing 
            nanobody technology for expression screening of membrane proteins.
  JOURNAL   Commun Biol. 2021 Mar 19;4(1):366. doi: 10.1038/s42003-021-01891-y.
  PUBMED    33742097
REFERENCE   2  (bases 1 to 226)
  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: "Commun 
            Biol."; date: "2021-03-19"; volume: "4(1):366. doi"; pages: " 
            10.1038/s42003-021-01891-y"
COMMENT     Sequence Label: pBMNb-cALFA
FEATURES             Location/Qualifiers
     source          1..226
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             29..52
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             62..100
                     /codon_start=1
                     /product="rationally designed epitope tag that forms a 
                     stable alpha-helix (Gotzke et al., 2019)"
                     /label=ALFA-tag
                     /note="It may be beneficial to flank the ALFA-tag sequence 
                     with proline residues to preserve the helical structure."
                     /translation="SRLEEELRRRLTE"
     CDS             join(110..133,134..169,170..193)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="WSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             200..223
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
ORIGIN
        1 gaattcaaag gcctacgtcg acggcagcct ggaggtgctg ttccagggcc ccagcggcgg
       61 gagccgtctc gaggaggagc tgagaagaag actgaccgaa ggcggcagct ggagccatcc
      121 ccagttcgaa aaaggcgggg gctccggtgg tggaagcggg ggatctgcgt ggagccaccc
      181 gcagttcgag aagggcagcc accaccacca ccaccaccac cactaa
//
