LOCUS       sequence-150052-         704 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression flag-GFP 11 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 704)
  AUTHORS   Lee JG, Takahama S, Zhang G, Tomarev SI, Ye Y
  TITLE     Unconventional secretion of misfolded proteins promotes adaptation 
            to proteasome dysfunction in mammalian cells.
  JOURNAL   Nat Cell Biol. 2016 Jul;18(7):765-76. doi: 10.1038/ncb3372. Epub 
            2016 Jun 13.
  PUBMED    27295555
REFERENCE   2  (bases 1 to 704)
  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"; doi: "10.1038/ncb3372"; 
            journalName: "Nat Cell Biol"; date: "2016-07"; volume: "18"; issue: 
            "7"; pages: "765-76"
COMMENT     Sequence Label: pRK-flag-GFP11
FEATURES             Location/Qualifiers
     source          1..704
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     43..60
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        80..98
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             complement(136..183)
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             complement(220..243)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      243..252
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(292..311)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(324..342)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(552..576)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(573..672)
                     /label=CMVd1 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate 
                     early promoter that yields slightly reduced expression 
                     relative to the full-length promoter"
     primer_bind     complement(602..622)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg
       61 ctgatcagcg agctctagca tttaggtgac actatagaat agggccctct agatgcatgc
      121 tcgagcggcc gcttatgtga tgccggcggc gttcacgtac tcgtgcagca ccatgtggtc
      181 ccggctgccg ccgccgctgc cgccgtcgcc ggtcgacccc ttgtcatcgt cgtccttgta
      241 gtccatggcg gcaagcttag aaccgaggtg cagttggacc tgggagtgga cacctgtgga
      301 gagaaaggca aagtggatgt tattctatag tgtcacctaa atcgtatgtg tatgatacat
      361 aaggttatgt attaattgta gccgcgttct aacgaagcca agggggtggg cctatagact
      421 ctataggcgg tacttacgtc actcttggca cggggaatcc gcgttccaat gcaccgttcc
      481 cggccgcgga ggctggatcg gtcccggtgt cttctatgga ggtcaaaaca gcgtggatgg
      541 cgtctccagg cgatctgacg gttcactaaa cgagctctgc ttatatagac ctcccaccgt
      601 acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt tggaaagtcc
      661 cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggg
//
