LOCUS       sequence-131897-         711 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Co-expressed human optimized S. pyogenese dCas9 fused to KRAB 
            repressor domain and GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 711)
  AUTHORS   Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
  TITLE     Multiplex CRISPR/Cas9-based genome engineering from a single 
            lentiviral vector.
  JOURNAL   Nucleic Acids Res. 2014 Aug 13. pii: gku749.
  PUBMED    25122746
REFERENCE   2  (bases 1 to 711)
  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: "Nucleic 
            Acids Res. 2014 Aug 13. pii: gku749."
COMMENT     Sequence Label: pLV hUbC-dCas9 KRAB-T2A-GFP
FEATURES             Location/Qualifiers
     source          1..711
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(13..66)
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             complement(73..93)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(160..354)
                     /codon_start=1
                     /product="Kruppel-associated box (KRAB) transcriptional 
                     repression domain from the human zinc finger protein ZNF10 
                     (Margolin et al., 1994)"
                     /label=KRAB
                     /translation="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             complement(388..408)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gctcacggta ccagggccgg gattctcctc cacgtcaccg catgttagaa gacttcctct
       61 gccctcgcta gcaactttgc gtttcttttt cggaactgat gatttgattt caaatgcagt
      121 ctctgaatca ggatgggtct cttggtgaat ttctctctcc accagccagg gctcttctcc
      181 cttctccaac cggaggatca catctggctt agtaagctga taacccaagg aaaccaggtt
      241 cttatagttc tccagcatca catttctgta caggatctgc tgagcagtgt ccagcagctt
      301 ccactcctcc ctggtgaagt ccacaaacac atccttgaag gtcaccagtg tccgggacca
      361 ggcagtcagt gacttagcat cgctagccac cttcctcttc ttcttggggt cagccctgct
      421 gtctccaccg agctgagaga ggtcgattct tgtttcatag agccccgtaa ttgactgatg
      481 aatcagtgtg gcgtccagga cctcctttgt agaggtgtac cgctttctgt ctatggtggt
      541 gtcgaagtac ttgaaggctg caggcgcgcc caagttggtc agagtaaaca agtggataat
      601 gttttctgcc tgctccctga tgggcttatc cctgtgctta ttgtaagcag aaagcacctt
      661 atcgaggtta gcgtcggcga ggatcactct tttggagaat tcgcttattt g
//
