LOCUS       sequence-109275-         767 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 767)
  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 767)
  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..767
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     65..87
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     80..96
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    112..211
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     213..229
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     239..258
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        239..257
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      293..302
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             311..331
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct agcatggatg
       61 ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc ccaaataatg
      121 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca atgctttttt
      181 ataatgccaa ctttgtacaa aaaagcaggc ttgtaaaacg acggccagtg agcgcgcgta
      241 atacgactca ctatagggcg aattgggtac cgggcccccc ctcgaggtcg acgccaccat
      301 ggcttctcca cctaagaaga agagaaaggt gggaagcatg gataagaagt atagcatcgg
      361 cctggatatt ggaactaact ccgtgggttg ggcagtgatt acagacgact acaaggtccc
      421 tagcaagaaa tttaaggtgc tgggtaacac cgacaggcac agcatcaaga aaaatctgat
      481 tggagccctg ctgttcggtt ctggagagac tgccgaagca acacgcctga aaagaacagc
      541 aagaaggcgc tataccagaa ggaagaatag aatctgttac ctgcaggaga ttttctctaa
      601 cgaaatggct aaggtggacg attcattctt tcataggctg gaggaaagtt tcctggtcga
      661 ggaagataag aaacacgagc gccatcctat ctttggaaac attgtggacg aggtcgccta
      721 tcacgaaaaa tacccaacca tctatcatct gcgcaagaaa ctggctg
//
