LOCUS Exported 598 bp ds-DNA linear SYN 18-MAY-2021 DEFINITION Expresses Cas9 nuclease and the PITCh-gRNA. ACCESSION . VERSION . KEYWORDS pX330S-2-PITCh SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 598) AUTHORS Sakuma T, Nakade S, Sakane Y, Suzuki KT, Yamamoto T TITLE MMEJ-assisted gene knock-in using TALENs and CRISPR-Cas9 with the PITCh systems. JOURNAL Nat Protoc. 2016 Jan;11(1):118-33. doi: 10.1038/nprot.2015.140. Epub 2015 Dec 17. PUBMED 26678082 REFERENCE 2 (bases 1 to 598) AUTHORS . TITLE Direct Submission JOURNAL Exported May 18, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..598 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(311..331) /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS complement(338..403) /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" regulatory 403..412 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" ORIGIN 1 agctgtcgtc caccttggcc atctcgttgc tgaagatctc ttgcagatag cagatccggt 61 tcttccgtct ggtgtatctt cttctggcgg ttctcttcag ccgggtggcc tcggctgttt 121 cgccgctgtc gaacagcagg gctccgatca ggttcttctt gatgctgtgc cggtcggtgt 181 tgcccagcac cttgaatttc ttgctgggca ccttgtactc gtcggtgatc acggcccagc 241 ccacagagtt ggtgccgatg tccaggccga tgctgtactt cttgtcggct gctgggactc 301 cgtggatacc gaccttccgc ttcttctttg gggccatctt atcgtcatcg tctttgtaat 361 caatatcatg atccttgtag tctccgtcgt ggtccttata gtccatggtg gcaccggtcc 421 aacctgaaaa aaagtgattt caggcaggtg ctccaggtaa ttaaacatta ataccccacc 481 aaccaaccat cccttaaacc cttacctctt gctcagctaa ttacagcccg gaggagaagg 541 gccgtcccgc ccgctcacct gtgggagtaa cgcggtcagt cagagccggg gcgggcgg //