LOCUS       sequence-78563-e         684 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 with mutated Ser and Thr residues in the 
            N-terminus and mutations at P676C I730C. Contains N-terminal 
            3xFLAG-YFP tag for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 684)
  AUTHORS   Monette MY, Somasekharan S, Forbush B
  TITLE     Molecular Motions Involved in Na-K-Cl Cotransporter-mediated Ion 
            Transport and Transporter Activation Revealed by Internal 
            Crosslinking between Transmembrane Domains 10 and 11/12.
  JOURNAL   J Biol Chem. 2014 Jan 22.
  PUBMED    24451383
REFERENCE   2  (bases 1 to 684)
  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. 2014 Jan 22."
COMMENT     Sequence Label: pcDNA3.1 Flag YFP hNKCC1 N-terminus T,S mutati...
FEATURES             Location/Qualifiers
     source          1..684
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     50..69
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      100..109
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             112..177
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(232..253)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(493..512)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
       61 actataggga gacccaagct ggctagcccg ggcacggccg ccaccatggg agactacaaa
      121 gaccatgacg gtgattataa agatcacgac atcgactaca aggatgacga tgacaagggt
      181 gaccctaccg tcagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      241 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      301 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      361 cccaccctcg tgaccacctt cggctacggc ctgatgtgct tcgcccgcta ccccgaccac
      421 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      481 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      541 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatccta
      601 gggcacaagc tggagtacaa ctacaacagc cacaacgttt atatcatggc cgacaagcag
      661 aagaacggca tcaaggtgaa cttc
//
