LOCUS       sequence-223229-        1411 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPRi negative control. KRAB domain and catalytically inactive 
            Cas9 from S. pyogenes with 2A-BlastR under the hPGK promoter, and 
            sgRNA2 against the EGFP CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1411)
  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 1411)
  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 EGFP-guide2
FEATURES             Location/Qualifiers
     source          1..1411
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(47..232)
                     /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      259..268
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(281..791)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(344..362)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_RNA        complement(822..897)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(977..996)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 annnnnatnc tgtanttctt gtnntcgacc ctccgccgga accgccgggc tcttctccct
       61 tctccaaccg gaggatcaca tctggcttag taagctgata acccaaggaa accaggttct
      121 tatagttctc cagcatcaca tttctgtaca cgatctgctg agcagtgtcc agcagcttcc
      181 actcctccct ggtgaagtcc acaaatacat ccttgaaggt caccagtgtc cgggaccagg
      241 cagttagtga cttagcatcc atggtggcgg atcaccggtc ctggggagag aggtcggtga
      301 ttcggtcaac gagggagccg actgccgacg tgcgctccgg aggcttgcag aatgcggaac
      361 accgcgcggg caggaacagg gcccacacta ccgccccaca ccccgcctcc cgcaccgccc
      421 cttcccggcc gctgctctcg gcgcgccctg ctgagcagcc gctattggcc acagcccatc
      481 gcggtcggcg cgctgccatt gctccctggc gctgtccgtc tgcgagggta ctagtgagac
      541 gtgcggcttc cgtttgtcac gtccggcacg ccgcgaaccg caaggaacct tcccgactta
      601 ggggcggagc aggaagcgtc gccggggggc ccacaagggt agcggcgaag atccgggtga
      661 cgctgcgaac ggacgtgaag aatgtgcgag acccagggtc ggcgccgctg cgtttcccgg
      721 aaccacgccc agagcagccg cgtccctgcg caaacccagg gctgccttgg aaaaggcgca
      781 accccaaccc ctttcaagac ctagctagcg aattcaaaaa agcaccgact cggtgccact
      841 ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct ctaaaacttt
      901 acgtcgccgt ccagctccgg tgtttcgtcc tttccacaag atatataaag ccaagaaatc
      961 gaaatacttt caagttacgg taagcatatg atagtccatt ttaaaacata attttaagac
     1021 tgcaaactac ccaagaaatt attactttct acgtcacgta ttttgtacaa aaatctttga
     1081 gtttacaggc aaattaattc gaattatccc cctaacaggc ctggaatcgg aaatgcaaaa
     1141 atggaaggaa tcatggggaa ataggccccc cggaacctta attaaccaaa ctgggattcc
     1201 cggctggccc cgggaataaa accccagaaa ttttggaaat tcttggaaat tcggttttag
     1261 gaaaatccgt taaagttgga gtaagccggt ttncanntat nnggtcnacc aatttctttt
     1321 gcccnagaac gtgganaacg ccaaaagacc ccncttaatt tgtttaaaaa ttggggaata
     1381 ggaaattgcn ccgtttgtct cgcaaaaaat a
//
