LOCUS       Exported                 862 bp ds-DNA     linear   SYN 11-JUN-2021
DEFINITION  Expresses human NKCC1 Y739C mutant with an N-terminal 3xFLAG-YFP tag
            in mammalian cells. cDNA is synthetic, containing convenient 
            restriction sites..
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1 Flag YFP hNKCC1 Y739C (NT525)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 862)
  TITLE     Forbush Lab human NKCC1 NT clones
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 862)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 11, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..862
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     50..69
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      100..109
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             112..177
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             193..705
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGLMCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIE"
     primer_bind     complement(232..253)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(493..512)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     840..861
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
       61 actataggga gacccaagct ggctagcccg ggcacggccg ccaccatggg agactacaaa
      121 gaccatgacg gtgattataa agatcacgac atcgactaca aggatgacga tgacaagggt
      181 gaccctaccg tcagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      241 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      301 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      361 cccaccctcg tgaccacctt cggctacggc ctgatgtgct tcgcccgcta ccccgaccac
      421 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      481 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      541 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatccta
      601 gggcacaagc tggagtacaa ctacaacagc cacaacgttt atatcatggc cgacaagcag
      661 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
      721 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      781 aaccactacc tgagctacca gtccaaactg agcaaagacc ccaacgagaa gcgcgatcac
      841 atggtcctgc tggagttcgt ga
//
