LOCUS       sequence-108209-         743 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Middle Entry clone for Gateway containing a zebrafish 
            codon-optimized Cas9 flanked by 2 NLS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 743)
  AUTHORS   Ablain J, Durand EM, Yang S, Zhou Y, Zon LI
  TITLE     A CRISPR/Cas9 Vector System for Tissue-Specific Gene Disruption in 
            Zebrafish.
  JOURNAL   Dev Cell. 2015 Mar 4. pii: S1534-5807(15)00075-1. doi: 
            10.1016/j.devcel.2015.01.032.
  PUBMED    25752963
REFERENCE   2  (bases 1 to 743)
  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: "Dev Cell. 
            2015 Mar 4. pii: S1534-5807(15)00075-1. doi: 
            10.1016/j.devcel.2015.01.032."
COMMENT     Sequence Label: pME-Cas9
FEATURES             Location/Qualifiers
     source          1..743
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(32..52)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     RBS             147..155
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
ORIGIN
        1 gcggtggcgg ccgctctaga ttccgcggtc acacctttct cttcttctta ggagatctca
       61 ccggtgagtc gccacccagc tgagacaggt caatccttgt ttcgtacagg ccggtaatac
      121 tctgatggat cagtgtagcg tccagcacct ccttagtgga ggtgtacctc tttctatcga
      181 ttgtagtgtc gaagtacttg aaagctgcag gtgcgcccag gtttgtcaga gtgaacagat
      241 gaatgatatt ttcggcctgc tccctgatgg gcttatctct gtgtttgttg taagcagaca
      301 ggaccttatc cagatttgcg tcggccagga tcactctttt agagaactcg ctaatctgtt
      361 caatgatctc gtccagataa tgcttgtgct gctccacaaa cagctgtttc tgttcgttat
      421 cctctggaga tcctttcagc ttctcgtagt ggcttgccag atacaggaag ttcacgtact
      481 tagaaggcag ggccagttca ttgcctttct gcagctctcc agcgcttgcc agcatgcgtt
      541 tcctgccgtt ttccagctca aacagactgt acttgggcag tttaatgatc aggtctttct
      601 tcacttcctt ataacctttg gcctccagaa aatcgattgg gttcttctcg aaagagcttc
      661 tctccataat tgtgataccc agcagctcct tgacagattt cagtttcttg cttttgccct
      721 tctccacctt agcgaccacc agg
//
