LOCUS       sequence-78728-e         742 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 S679C I730C 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 742)
  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 742)
  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 S679C I730C (NT838)
FEATURES             Location/Qualifiers
     source          1..742
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     52..71
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        52..70
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      102..111
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             114..179
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(192..194,195..707)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIE"
     primer_bind     complement(234..255)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(495..514)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
       61 tcactatagg gagacccaag ctggctagcc cgggcacggc cgccaccatg ggagactaca
      121 aagaccatga cggtgattat aaagatcacg acatcgacta caaggatgac gatgacaagg
      181 gtgaccctac cgtcagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      241 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      301 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      361 ggcccaccct cgtgaccacc ttcggctacg gcctgatgtg cttcgcccgc taccccgacc
      421 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      481 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      541 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      601 tagggcacaa gctggagtac aactacaaca gccacaacgt ttatatcatg gccgacaagc
      661 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      721 agctcgccga ccactaccag ca
//
