LOCUS       sequence-129980-         896 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express sgRNA and dCas9-KRAB from 3rd generation lentiviral vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 896)
  AUTHORS   Thakore PI, D'Ippolito AM, Song L, Safi A, Shivakumar NK, Kabadi AM,
            Reddy TE, Crawford GE, Gersbach CA
  TITLE     Highly specific epigenome editing by CRISPR-Cas9 repressors for 
            silencing of distal regulatory elements.
  JOURNAL   Nat Methods. 2015 Dec;12(12):1143-9. doi: 10.1038/nmeth.3630. Epub 
            2015 Oct 26.
  PUBMED    26501517
REFERENCE   2  (bases 1 to 896)
  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"; doi: 
            "10.1038/nmeth.3630"; journalName: "Nat Methods"; date: "2015-12"; 
            volume: "12"; issue: "12"; pages: "1143-9"
COMMENT     Sequence Label: pLV hU6-sgRNA hUbC-dCas9-KRAB-T2a-GFP
FEATURES             Location/Qualifiers
     source          1..896
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(16..69)
                     /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(76..96)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(163..357)
                     /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(391..411)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 cttgctcacg gtaccagggc cgggattctc ctccacgtca ccgcatgtta gaagacttcc
       61 tctgccctcg ctagcaactt tgcgtttctt tttcggaact gatgatttga tttcaaatgc
      121 agtctctgaa tcaggatggg tctcttggtg aatttctctc tccaccagcc agggctcttc
      181 tcccttctcc aaccggagga tcacatctgg cttagtaagc tgataaccca aggaaaccag
      241 gttcttatag ttctccagca tcacatttct gtacaggatc tgctgagcag tgtccagcag
      301 cttccactcc tccctggtga agtccacaaa cacatccttg aaggtcacca gtgtccggga
      361 ccaggcagtc agtgacttag catcgctagc caccttcctc ttcttcttgg ggtcagccct
      421 gctgtctcca ccgagctgag agaggtcgat tcttgtttca tagagccccg taattgactg
      481 atgaatcagt gtggcgtcca ggacctcctt tgtagaggtg taccgctttc tgtctatggt
      541 ggtgtcgaag tacttgaagg ctgcaggcgc gcccaagttg gtcagagtaa acaagtggat
      601 aatgttttct gcctgctccc tgatgggctt atccctgtgc ttattgtaag cagaaagcac
      661 cttatcgagg ttagcgtcgg cgaggatcac tcttttggag aattcgctta tttgctcgat
      721 gatctcatca aggtagtgtt tgtgttgttc cacgaacagc tgcttctgct cattatcttc
      781 gggagaccct ttgagctttt catagtggct ggccagatac aagaaattaa cgtatttaga
      841 gggcagtgcc agctcgttac ctttctgcag ctcgcccgca ctagcgagca ttcgtt
//
