LOCUS       sequence-354713-         653 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For expression of membrane protein with N-terminal FLAG tag and 
            C-terminal split GFP11 fragment.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 653)
  AUTHORS   Bruguera ES, Mahoney JP, Weis WI
  TITLE     Reconstitution of purified membrane protein dimers in lipid 
            nanodiscs with defined stoichiometry and orientation using a split 
            GFP tether.
  JOURNAL   J Biol Chem. 2022 Jan 21:101628. doi: 10.1016/j.jbc.2022.101628.
  PUBMED    35074428
REFERENCE   2  (bases 1 to 653)
  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: "J Biol 
            Chem. 2022 Jan 21:101628. doi: 10.1016/j.jbc.2022.101628."
COMMENT     Sequence Label: FLAG-3C-sGFP11_pcDNA3.1(-)
FEATURES             Location/Qualifiers
     source          1..653
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     23..42
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        23..41
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             184..207
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             220..267
                     /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"
     primer_bind     complement(303..320)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    309..533
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
       61 gtttaaacgg gccctctaga ctcgagcggc cgcgccacca tgaagacgat catcgccctg
      121 agctacatct tctgcctggt gttcgccgac tacaaggacg atgatgacgc caccggtgga
      181 tccctcgaag tccttttcca gggccctggt tctggttctc gtgaccacat ggtgctgcat
      241 gaatatgtga acgcggcggg catcacgtga ggtaccaagc ttaagtttaa accgctgatc
      301 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
      361 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
      421 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
      481 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
      541 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
      601 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcg
//
