LOCUS       sequence-145837-         725 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Ubiquitin promoter expresses GFP and humanized spCas9 (PX330) via a 
            T2A motif. For cell transfection or use in retroviral (MMuLV) 
            packaging. Use Pac1 and/or BstB1 sites to insert sgRNAs from PXL..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 725)
  AUTHORS   Williams MR, Fricano-Kugler CJ, Getz SA, Skelton PD, Lee J, Rizzuto 
            CP, Geller JS, Li M, Luikart BW
  TITLE     A Retroviral CRISPR-Cas9 System for Cellular Autism-Associated 
            Phenotype Discovery in Developing Neurons.
  JOURNAL   Sci Rep. 2016 May 10;6:25611. doi: 10.1038/srep25611.
  PUBMED    27161796
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: "Sci Rep."; 
            date: "2016-05-10"; volume: "6:25611. doi"; pages: " 
            10.1038/srep25611"
COMMENT     Sequence Label: pRubiG-T2A-Cas9
FEATURES             Location/Qualifiers
     source          1..725
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      161..170
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(167..169,170..172,173..685)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(212..233)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(473..492)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 gtgagttttt aggcaccttt tgaatgtaat catttgggtc aatatgtaat tttcagtgtt
       61 agactagtaa attgtccgct aaattctggc cgtttttggc ttttttgtta gacgaagctt
      121 gggctgcagg tcgactctag aggatccccg ggtaccggtc gccaccatgg tgagcaaggg
      181 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
      241 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
      301 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
      361 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
      421 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
      481 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
      541 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
      601 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
      661 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
      721 gaaca
//
