LOCUS       sequence-108212-         872 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 872)
  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 872)
  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..872
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    74..98
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     99..116
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     100..116
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     126..145
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        126..144
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      180..189
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             198..218
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aataatgatt ttattttgac tgatagtgac ctgttcgttg caacaaattg atgagcaatg
       61 cttttttata atgccaactt tgtacaaaaa agcaggcttg taaaacgacg gccagtgagc
      121 gcgcgtaata cgactcacta tagggcgaat tgggtaccgg gccccccctc gaggtcgacg
      181 ccaccatggc ttctccacct aagaagaaga gaaaggtggg aagcatggat aagaagtata
      241 gcatcggcct ggatattgga actaactccg tgggttgggc agtgattaca gacgactaca
      301 aggtccctag caagaaattt aaggtgctgg gtaacaccga caggcacagc atcaagaaaa
      361 atctgattgg agccctgctg ttcggttctg gagagactgc cgaagcaaca cgcctgaaaa
      421 gaacagcaag aaggcgctat accagaagga agaatagaat ctgttacctg caggagattt
      481 tctctaacga aatggctaag gtggacgatt cattctttca taggctggag gaaagtttcc
      541 tggtcgagga agataagaaa cacgagcgcc atcctatctt tggaaacatt gtggacgagg
      601 tcgcctatca cgaaaaatac ccaaccatct atcatctgcg caagaaactg gctgactcta
      661 ctgataaagc cgacctgaga ctgatctatc tggctctggc ccacatgatt aagttcaggg
      721 gtcattttct gatcgagggc gatctgaacc ccgacaattc cgatgtggac aagctgttca
      781 tccagctggt ccagatttac aatcagctgt ttgaggaaaa ccctattaat gcttccagag
      841 tggacgcaaa agctatcctg tcagccaggc tg
//
