LOCUS       sequence-87770-e         725 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Csy4 and Cas9 D10A nickase expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 725)
  AUTHORS   Tsai SQ, Wyvekens N, Khayter C, Foden JA, Thapar V, Reyon D, Goodwin
            MJ, Aryee MJ, Joung JK
  TITLE     Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome 
            editing.
  JOURNAL   Nat Biotechnol. 2014 Apr 25. doi: 10.1038/nbt.2908.
  PUBMED    24770325
REFERENCE   2  (bases 1 to 725)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2014 Apr 25. doi: 10.1038/nbt.2908."
COMMENT     Sequence Label: pNW3
FEATURES             Location/Qualifiers
     source          1..725
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      41..50
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             47..607
                     /codon_start=1
                     /product="CRISPR-associated endoribonuclease responsible 
                     for pre-crRNA processing in Pseudomonas aeruginosa 
                     (Haurwitz et al., 2010)"
                     /label=Csy4
                     /translation="MDHYLDIRLRPDPEFPPAQLMSVLFGKLHQALVAQGGDRIGVSFP
                     DLDESRSRLGERLRIHASADDLRALLARPWLEGLRDHLQFGEPAVVPHPTPYRQVSRVQ
                     AKSNPERLRRRLMRRHDLSEEEARKRIPDTVARALDLPFVTLRSQSTGQHFRLFIRHGP
                     LQVTAEEGGFTCYGLSKGGFVPWF"
     CDS             608..661
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
ORIGIN
        1 tctgcagtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg accactacct
       61 cgacattcgc ttgcgaccgg acccggaatt tcccccggcg caactcatga gcgtgctctt
      121 cggcaagctc caccaggccc tggtggcaca gggcggggac aggatcggcg tgagcttccc
      181 cgacctcgac gaaagccgct cccggctggg cgagcgcctg cgcattcatg cctcggcgga
      241 cgaccttcgt gccctgctcg cccggccctg gctggaaggg ttgcgggacc atctgcaatt
      301 cggagaaccg gcagtcgtgc ctcaccccac accgtaccgt caggtcagtc gggttcaggc
      361 gaaaagcaat ccggaacgcc tgcggcggcg gctcatgcgc cggcacgatc tgagtgagga
      421 ggaggctcgg aaacgcattc ccgatacggt cgcgagagcc ttggacctgc ccttcgtcac
      481 gctacgcagc cagagcaccg gacagcactt ccgtctcttc atccgccacg ggccgttgca
      541 ggtgacggca gaggaaggag gattcacctg ttacgggttg agcaaaggag gtttcgttcc
      601 ctggttcgag ggcagaggaa gtctgttaac atgcggtgac gtcgaggaga atcctggccc
      661 aatggataaa aagtattcta ttggtttagc catcggcact aattccgttg gatgggctgt
      721 cataa
//
