LOCUS Exported 746 bp ds-DNA linear SYN 08-JUL-2021 DEFINITION Expresses human NKCC1 G719C C723S C724V 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 G719C C723S C724V (NT533) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 746) TITLE Forbush Lab human NKCC1 NT clones JOURNAL Unpublished REFERENCE 2 (bases 1 to 746) AUTHORS . TITLE Direct Submission JOURNAL Exported Jul 8, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..746 /organism="synthetic DNA construct" /mol_type="other DNA" 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 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 109..174 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 190..702 /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(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 catcctaggg 601 cacaagctgg agtacaacta caacagccac aacgtttata tcatggccga caagcagaag 661 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc 721 gccgaccact accagcagaa cacccc //