LOCUS       sequence-79783-e         989 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 with truncation of N-terminus, mutations at 
            P676C I730C and C-terminus replaced with that of NBCe. Contains an 
            N-terminal 3xFLAG-YFP tag for expression in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 989)
  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 989)
  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 N-termina...
FEATURES             Location/Qualifiers
     source          1..989
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      15..24
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             27..92
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             join(105..107,108..818)
                     /codon_start=1
                     /product="enhanced variant of YFP (Heikal et al., 2001)"
                     /label=Citrine
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(105..107,108..818)
                     /codon_start=1
                     /product="enhanced YFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEYFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(147..168)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(408..427)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     755..776
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gcccgggcac ggccgccacc atgggagact acaaagacca tgacggtgat tataaagatc
       61 acgacatcga ctacaaggat gacgatgaca agggtgaccc taccgtcagc aagggcgagg
      121 agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca
      181 agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt
      241 tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accttcggct
      301 acggcctgat gtgcttcgcc cgctaccccg accacatgaa gcagcacgac ttcttcaagt
      361 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      421 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      481 agggcatcga cttcaaggag gacggcaaca tcctagggca caagctggag tacaactaca
      541 acagccacaa cgtttatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
      601 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
      661 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc taccagtcca
      721 aactgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
      781 ccgccgggat cactctcggc atggacgagc tctacaaggg tgacctggag ccgaggccca
      841 cggctcctag ctccggcgcg cgcgccagga tcgaccacta ccggcacaca gccgcgcagc
      901 tcggcgagaa gctgctccgg cctagcctgg ccgagctgca tgacgagctg gaaaaggaac
      961 cttttgagga tggctttgca aatggggaa
//
