LOCUS       sequence-79779-e         990 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses human NKCC1 L722C C723S C724V 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 990)
  TITLE     Forbush Lab human NKCC1 NT clones
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 990)
  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            "Unpublished"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 L722C C723S C724V (NT...
FEATURES             Location/Qualifiers
     source          1..990
                     /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 ccaggcctcg ccggagtcgg cgaaacgccg agcgccgctg
      901 cgctggccgc agccagggtg gaactgcccg gaaccgctgt gccctcggtg ccggaggatg
      961 ctgcgcccgc gagccgggac gggggcgggg
//
