LOCUS       sequence-78578-e         655 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 C723S C724V F728C mutant with an N-terminal 
            3xFLAG-YFP tag in mammalian cells. cDNA is synthetic, containing 
            convenient restriction sites..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 655)
  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 655)
  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 C723S C724V F728C (NT...
FEATURES             Location/Qualifiers
     source          1..655
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     49..68
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        49..67
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      99..108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             111..176
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(231..252)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(492..511)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
       61 ctatagggag acccaagctg gctagcccgg gcacggccgc caccatggga gactacaaag
      121 accatgacgg tgattataaa gatcacgaca tcgactacaa ggatgacgat gacaagggtg
      181 accctaccgt cagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      241 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      301 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      361 ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt cgcccgctac cccgaccaca
      421 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      481 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      541 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctag
      601 ggcacaagct ggagtacaac tacaacagcc acaacgttta tatcatggcc gacaa
//
