LOCUS       sequence-148127-         951 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Tet-regulatable dCas9-KRAB 2nd generation lentiviral expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 951)
  AUTHORS   Kearns NA, Genga RM, Enuameh MS, Garber M, Wolfe SA, Maehr R
  TITLE     Cas9 effector-mediated regulation of transcription and 
            differentiation in human pluripotent stem cells.
  JOURNAL   Development. 2014 Jan;141(1):219-23. doi: 10.1242/dev.103341.
  PUBMED    24346702
REFERENCE   2  (bases 1 to 951)
  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: "10.1242/dev"; 
            journalName: "Development"; date: "2014-01"; volume: "141"; issue: 
            "1"; pages: "219-23"
COMMENT     Sequence Label: pHAGE TRE dCas9-KRAB
FEATURES             Location/Qualifiers
     source          1..951
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(182..208)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     protein_bind    261..285
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(326..520)
                     /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="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
                     QLTKPDVILRLEKGEEPWLV"
     CDS             complement(557..646)
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             complement(647..667)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gatcaggacg ctcgctgcgc ccttcgtctg acgtggcagc gctcgccgtg aggagggggg
       61 cgcccgcggg aggcgccaaa acccggcgcg gaggccagat cttgggtggg ttactccaga
      121 ctgccttggg aaaagcgcct cccctacccg gtagaatttc tagtttaatt aatcattact
      181 aagcgtagtc tgggacgtcg tatgggtatt cgaaccgcgg gccctctaga ctcgagcggc
      241 cgccactgtg ctggatatca accactttgt acaagaaagc tgggtctaga tatctcgagg
      301 cggccgctta tccggattcg aattatacca gccaaggttc ttcccccttt tcgagcctaa
      361 ggatcacatc cggtttcgtc aactggtatc cgagactgac gaggttcttg taattctcaa
      421 gcatgacatt tcgatacaca atttgttgcg cagtatccag caatttccac tcctcacgtg
      481 taaagtccac aaacacgtcc ttgaacgtaa ccagtgttct cgaccatgcc gtaagtgatt
      541 tcgcgtccat ggatccagcg taatctggaa cgtcataagg atacgatcct gcatagtccg
      601 ggacgtcata gggatagccc gcatagtcag gaacatcgta tgggtatacc ttcctcttct
      661 tcttgggccc gccggtgccg tctccaccga gctgagagag gtcgattctt gtttcataga
      721 gccccgtaat tgactgatga atcagtgtgg cgtccaggac ctcctttgta gaggtgtacc
      781 gctttctgtc tatggtggtg tcgaagtact tgaaggctgc aggcgcgccc aagttggtca
      841 gagtaaacaa gtggataatg ttttctgcct gctccctgat gggcttatcc ctgtgcttat
      901 tgtaagcaga aagcacctta tcgaggttag cgtcggcgag gatcactctt t
//
