LOCUS       sequence-78729-e         597 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 V729C 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 597)
  TITLE     Forbush Lab human NKCC1 NT clones
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 597)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pcDNA3.1 Flag YFP hNKCC1 V729C (NT503)
FEATURES             Location/Qualifiers
     source          1..597
                     /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"
     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"
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 catccta
//
