LOCUS       sequence-78616-e         718 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 W732C 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 718)
  TITLE     Forbush Lab human NKCC1 NT clones
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 718)
  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            "Unpublished"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 W732C (NT506)
FEATURES             Location/Qualifiers
     source          1..718
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     49..68
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        49..67
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      99..108
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             111..176
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(189..191,192..704)
                     /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(231..252)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(492..511)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
       61 ctatagggag acccaagctg gctagcccgg gcacggccgc caccatggga gactacaaag
      121 accatgacgg tgattataaa gatcacgaca tcgactacaa ggatgacgat gacaagggtg
      181 accctaccgt cagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      241 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      301 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      361 ccaccctcgt gaccaccttc ggctacggcc tgatgtgctt cgcccgctac cccgaccaca
      421 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      481 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      541 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctag
      601 ggcacaagct ggagtacaac tacaacagcc acaacgttta tatcatggcc gacaagcaga
      661 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgca
//
