LOCUS Exported 977 bp ds-DNA linear SYN 22-AUG-2022 DEFINITION Cas9 from S. pyogenes with 2A-Puro, and cloning backbone for sgRNA (V2.0). ACCESSION . VERSION . KEYWORDS pSpCas9(BB)-2A-Puro (PX459) V2.0 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 977) AUTHORS Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F TITLE Genome engineering using the CRISPR-Cas9 system. JOURNAL Nat Protoc. 2013 Nov;8(11):2281-308. doi: 10.1038/nprot.2013.143. Epub 2013 Oct 24. PUBMED 24157548 REFERENCE 2 (bases 1 to 977) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 22, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..977 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 17..26 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 26..91 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 98..118 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" ORIGIN 1 ttcaggttgg accggtgcca ccatggacta taaggaccac gacggagact acaaggatca 61 tgatattgat tacaaagacg atgacgataa gatggcccca aagaagaagc ggaaggtcgg 121 tatccacgga gtcccagcag ccgacaagaa gtacagcatc ggcctggaca tcggcaccaa 181 ctctgtgggc tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt 241 gctgggcaac accgaccggc acagcatcaa gaagaacctg atcggagccc tgctgttcga 301 cagcggcgaa acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag 361 acggaagaac cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga 421 cgacagcttc ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga 481 gcggcacccc atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac 541 catctaccac ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat 601 ctatctggcc ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct 661 gaaccccgac aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca 721 gctgttcgag gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc 781 cagactgagc aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa 841 gaatggcctg ttcggaaacc tgattgccct gagcctgggc ctgaccccca acttcaagag 901 caacttcgac ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga 961 cctggacaac ctgctgg //