LOCUS       sequence-145826-         811 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Ubiquitin promoter expresses GFP and humanized spCas9 (from PX330) 
            via a T2A motif. For cell transfection or use in lentiviral 
            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 811)
  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 811)
  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: FUG-T2A-Cas9
FEATURES             Location/Qualifiers
     source          1..811
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      137..146
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(143..145,146..148,149..661)
                     /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(188..209)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(449..468)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 tgtaatcatt tgggtcaata tgtaattttc agtgttagac tagtaaattg tccgctaaat
       61 tctggccgtt tttggctttt ttgttagacg aagcttgggc tgcaggtcga ctctagagga
      121 tccccgggta ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      181 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      241 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      301 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
      361 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      421 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      481 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      541 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
      601 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
      661 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
      721 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
      781 cgagaagcgc gatcacatgg tcctgctgga g
//
