LOCUS       sequence-73823-e         886 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 with an N-terminal 
            3xFlag-YFP-(chloride-sensing) tag in mammalian cells. cDNA is 
            synthetic, containing convenient restriction sites. Native hNKCC1 
            amino acid sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 886)
  AUTHORS   Somasekharan S, Monette MY, Forbush B
  TITLE     Functional Expression of Human NKCC1 from a Synthetic Cassette-Based
            cDNA: Introduction of Extracellular Epitope Tags and Removal of 
            Cysteines.
  JOURNAL   PLoS One. 2013 Dec 5;8(12):e82060. doi: 
            10.1371/journal.pone.0082060.
  PUBMED    24339991
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: "PLoS One.";
            date: "2013-12-5"; volume: "8(12):e82060. doi"; pages: " 
            10.1371/journal.pone.0082060"
COMMENT     Sequence Label: pcDNA3.1 Flag Cl-sensitive YFP hNKCC1 WT (NT13)
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
                     FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKS
                     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 cagtgcttcg cccgctaccc cgaccacatg
      421 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      481 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      541 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      601 cacaagctgg agtacaacta caacagccac aacgtttata tcatggccga caagcagaag
      661 aacggcatca agtcgaattt caagatccgc cacaacatcg aagacggcag cgtgcagctc
      721 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      781 cactacctga gctaccagtc caaactgagc aaagacccca acgagaagcg cgatcacatg
      841 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatgg
//
