LOCUS       sequence-79701-e         832 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 P676C 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 832)
  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 832)
  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 P676C I730C (NT809)
FEATURES             Location/Qualifiers
     source          1..832
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     57..76
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        57..75
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      107..116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             119..184
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(197..199,200..712)
                     /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(239..260)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(500..519)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ataagcagag ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat
       61 acgactcact atagggagac ccaagctggc tagcccgggc acggccgcca ccatgggaga
      121 ctacaaagac catgacggtg attataaaga tcacgacatc gactacaagg atgacgatga
      181 caagggtgac cctaccgtca gcaagggcga ggagctgttc accggggtgg tgcccatcct
      241 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      301 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      361 gccctggccc accctcgtga ccaccttcgg ctacggcctg atgtgcttcg cccgctaccc
      421 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      481 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      541 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      601 catcctaggg cacaagctgg agtacaacta caacagccac aacgtttata tcatggccga
      661 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
      721 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
      781 gcccgacaac cactacctga gctaccagtc caaactgagc aaagacccca ac
//
