LOCUS Exported 811 bp ds-DNA linear SYN 23-JUN-2021 DEFINITION Expresses human NKCC1 A734C 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 A734C (NT508) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 811) TITLE Forbush Lab human NKCC1 NT clones JOURNAL Unpublished REFERENCE 2 (bases 1 to 811) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 23, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..811 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 61..80 /label=T7 /note="T7 promoter, forward primer" promoter 61..79 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" regulatory 111..120 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 123..188 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 204..716 /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(243..264) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(504..523) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" ORIGIN 1 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat 61 taatacgact cactataggg agacccaagc tggctagccc gggcacggcc gccaccatgg 121 gagactacaa agaccatgac ggtgattata aagatcacga catcgactac aaggatgacg 181 atgacaaggg tgaccctacc gtcagcaagg gcgaggagct gttcaccggg gtggtgccca 241 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg 301 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc 361 ccgtgccctg gcccaccctc gtgaccacct tcggctacgg cctgatgtgc ttcgcccgct 421 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc 481 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt 541 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg 601 gcaacatcct agggcacaag ctggagtaca actacaacag ccacaacgtt tatatcatgg 661 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg 721 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc 781 tgctgcccga caaccactac ctgagctacc a //