LOCUS       sequence-223213-        1066 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Catalytically inactive Cas9 from S. pyogenes with P2A-BlastR under 
            the hPGK promoter, and cloning backbone for sgRNA. Contains BsmBI 
            sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1066)
  AUTHORS   Miguel-Escalada I, Bonas-Guarch S, Cebola I, Ponsa-Cobas J, 
            Mendieta-Esteban J, Atla G, Javierre BM, Rolando DMY, Farabella I, 
            Morgan CC, Garcia-Hurtado J, Beucher A, Moran I, Pasquali L, 
            Ramos-Rodriguez M, Appel EVR, Linneberg A, Gjesing AP, Witte DR, 
            Pedersen O, Grarup N, Ravassard P, Torrents D, Mercader JM, Piemonti
            L, Berney T, de Koning EJP, Kerr-Conte J, Pattou F, Fedko IO, Groop 
            L, Prokopenko I, Hansen T, Marti-Renom MA, Fraser P, Ferrer J
  TITLE     Human pancreatic islet three-dimensional chromatin architecture 
            provides insights into the genetics of type 2 diabetes.
  JOURNAL   Nat Genet. 2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi: 
            10.1038/s41588-019-0457-0.
  PUBMED    31253982
REFERENCE   2  (bases 1 to 1066)
  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: "Nat Genet. 
            2019 Jun 28. pii: 10.1038/s41588-019-0457-0. doi: 
            10.1038/s41588-019-0457-0."
COMMENT     Sequence Label: Lenti-(BB)-hPGK-KRAB-dCas9-P2A-BlastR
FEATURES             Location/Qualifiers
     source          1..1066
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(69..254)
                     /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
                     QLTKPDVILRLEKGEEP"
     regulatory      281..290
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(303..813)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(366..384)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_RNA        complement(844..919)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gannagttgg tgccgatggc caggccgatg ctgtacttct tgtccatcga ccctccgccg
       61 gaaccgccgg gctcttctcc cttctccaac cggaggatca catctggctt agtaagctga
      121 taacccaagg aaaccaggtt cttatagttc tccagcatca catttctgta cacgatctgc
      181 tgagcagtgt ccagcagctt ccactcctcc ctggtgaagt ccacaaatac atccttgaag
      241 gtcaccagtg tccgggacca ggcagttagt gacttagcat ccatggtggc ggatcaccgg
      301 tcctggggag agaggtcggt gattcggtca acgagggagc cgactgccga cgtgcgctcc
      361 ggaggcttgc agaatgcgga acaccgcgcg ggcaggaaca gggcccacac taccgcccca
      421 caccccgcct cccgcaccgc cccttcccgg ccgctgctct cggcgcgccc tgctgagcag
      481 ccgctattgg ccacagccca tcgcggtcgg cgcgctgcca ttgctccctg gcgctgtccg
      541 tctgcgaggg tactagtgag acgtgcggct tccgtttgtc acgtccggca cgccgcgaac
      601 cgcaaggaac cttcccgact taggggcgga gcaggaagcg tcgccggggg gcccacaagg
      661 gtagcggcga agatccgggt gacgctgcga acggacgtga agaatgtgcg agacccaggg
      721 tcggcgccgc tgcgtttccc ggaaccacgc ccagagcagc cgcgtccctg cgcaaaccca
      781 gggctgcctt ggaaaaggcg caaccccaac ccctttcaag acctagctag cgaattcaaa
      841 aaagcaccga ctcggtgcca ctttttcaag ttgataacgg actagcctta ttttaacttg
      901 ctatttctag ctctaaaaca gagacgaaac aaaaaaaagc aagaacccaa aaaagattta
      961 aaaattattt ttagcgcagt taatggaaca ggaactaaat tttaccccaa aaatattacg
     1021 ngnaatcagg aaataaggtt attgaggttn nnggaccnnn ccttng
//
