LOCUS Exported 891 bp ds-DNA linear SYN 19-MAY-2021 DEFINITION Encode hSpCas9 for in vitro transcription with Sp6. ACCESSION . VERSION . KEYWORDS pCS2+hSpCas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 891) AUTHORS Ansai S, Kinoshita M TITLE Targeted mutagenesis using CRISPR/Cas system in medaka. JOURNAL Biol Open. 2014 Apr 11;3(5):362-71. doi: 10.1242/bio.20148177. PUBMED 24728957 REFERENCE 2 (bases 1 to 891) AUTHORS . TITLE Direct Submission JOURNAL Exported May 19, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..891 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 37..55 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 37..54 /label=SP6 /note="SP6 promoter, forward primer" regulatory 87..96 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 96..161 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 168..188 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" ORIGIN 1 accgccattc tgcctgggga cgtcggagca agcttgattt aggtgacact atagaataca 61 agctacttgt tctttttgca ggatccgcca ccatggacta taaggaccac gacggagact 121 acaaggatca tgatattgat tacaaagacg atgacgataa gatggcccca aagaagaagc 181 ggaaggtcgg tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca 241 tcggcaccaa ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga 301 aattcaaggt gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc 361 tgctgttcga cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa 421 gatacaccag acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg 481 ccaaggtgga cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata 541 agaagcacga gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga 601 agtaccccac catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc 661 tgcggctgat ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg 721 agggcgacct gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga 781 cctacaacca gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca 841 tcctgtctgc cagactgagc aagagcagac ggctggaaaa tctgatcgcc c //