LOCUS       sequence-78683-e         886 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 C723S C724V A735C 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 886)
  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 886)
  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 A735C (NT...
FEATURES             Location/Qualifiers
     source          1..886
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     47..66
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        47..65
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      97..106
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             109..174
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(187..189,190..702)
                     /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(229..250)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(490..509)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     837..858
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
       61 atagggagac ccaagctggc tagcccgggc acggccgcca ccatgggaga ctacaaagac
      121 catgacggtg attataaaga tcacgacatc gactacaagg atgacgatga caagggtgac
      181 cctaccgtca gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      241 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      301 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      361 accctcgtga ccaccttcgg ctacggcctg atgtgcttcg cccgctaccc cgaccacatg
      421 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      481 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      541 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctaggg
      601 cacaagctgg agtacaacta caacagccac aacgtttata tcatggccga caagcagaag
      661 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
      721 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      781 cactacctga gctaccagtc caaactgagc aaagacccca acgagaagcg cgatcacatg
      841 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatgg
//
