LOCUS       sequence-223231-        1303 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 
            sgRNA3 against the EGFP CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1303)
  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 1303)
  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-guide3
FEATURES             Location/Qualifiers
     source          1..1303
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     28..47
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        127..202
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        233..743
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     662..680
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     regulatory      756..765
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             792..977
                     /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
                     QLTKPDVILRLEKG*EP"
ORIGIN
        1 ttgnagtttt aaattatgtt ttaaatggac tatcatatgc ttaccgtaac ttgaaagtat
       61 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccggatg ccgttcttct
      121 gcttgtgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      181 aaaagtggca ccgagtcggt gcttttttga attcgctagc taggtcttga aaggggttgg
      241 ggttgcgcct tttccaaggc agccctgggt ttgcgcaggg acgcggctgc tctgggcgtg
      301 gttccgggaa acgcagcggc gccgaccctg ggtctcgcac attcttcacg tccgttcgca
      361 gcgtcacccg gatcttcgcc gctacccttg tgggcccccc ggcgacgctt cctgctccgc
      421 ccctaagtcg ggaaggttcc ttgcggttcg cggcgtgccg gacgtgacaa acggaagccg
      481 cacgtctcac tagtaccctc gcagacggac agcgccaggg agcaatggca gcgcgccgac
      541 cgcgatgggc tgtggccaat agcggctgct cagcagggcg cgccgagagc agcggccggg
      601 aaggggcggt gcgggaggcg gggtgtgggg cggtagtgtg ggccctgttc ctgcccgcgc
      661 ggtgttccgc attctgcaag cctccggagc gcacgtcggc agtcggctcc ctcgttgacc
      721 gaatcaccga cctctctccc caggaccggt gatccgccac catggatgct aagtcactaa
      781 ctgcctggtc ccggacactg gtgaccttca aggatgtatt tgtggacttc accagggagg
      841 agtggaagct gctggacact gctcagcaga tcgtgtacag aaatgtgatg ctggagaact
      901 ataagaacct ggtttccttg ggttatcagc ttactaagcc agatgtgatc ctccggttgg
      961 agaagggata agagcccggc ggtttccggc ggagggtcga tggacaagaa gtacagcatg
     1021 cgcctggcca tcggcaacac ttctgtcggc tgggaccgtg atcaccgaac aagtacaagg
     1081 tgcccancaa gaaattcaag gtgctgggcc acntccaacc ggcaccagca tcaggaaaga
     1141 cctggatccg gccccctgcc gtttcaaaca ccgaaaaaac nagtccaagg ccacccngct
     1201 tgaataaaac ccccctaaca aaagaatccc ctaaagggaa taaccggaat tgactttttt
     1261 gcaaaaaatt cttcccggcg gaaaggttgc ccaaggggtg gcg
//
