LOCUS       sequence-223225-        1548 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 
            sgRNA1 against the BFP CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1548)
  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 1548)
  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 BFP-guide1
FEATURES             Location/Qualifiers
     source          1..1548
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(49..234)
                     /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      261..270
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(283..793)
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     primer_bind     complement(346..364)
                     /label=hPGK-F
                     /note="Human PGK promoter, forward primer"
     misc_RNA        complement(824..899)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(979..998)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 cnnnnnnnat nctgtanttc ttgtnatcga ccctccgccg gaaccgccgg gctcttctcc
       61 cttctccaac cggaggatca catctggctt agtaagctga taacccaagg aaaccaggtt
      121 cttatagttc tccagcatca catttctgta cacgatctgc tgagcagtgt ccagcagctt
      181 ccactcctcc ctggtgaagt ccacaaatac atccttgaag gtcaccagtg tccgggacca
      241 ggcagttagt gacttagcat ccatggtggc ggatcaccgg tcctggggag agaggtcggt
      301 gattcggtca acgagggagc cgactgccga cgtgcgctcc ggaggcttgc agaatgcgga
      361 acaccgcgcg ggcaggaaca gggcccacac taccgcccca caccccgcct cccgcaccgc
      421 cccttcccgg ccgctgctct cggcgcgccc tgctgagcag ccgctattgg ccacagccca
      481 tcgcggtcgg cgcgctgcca ttgctccctg gcgctgtccg tctgcgaggg tactagtgag
      541 acgtgcggct tccgtttgtc acgtccggca cgccgcgaac cgcaaggaac cttcccgact
      601 taggggcgga gcaggaagcg tcgccggggg gcccacaagg gtagcggcga agatccgggt
      661 gacgctgcga acggacgtga agaatgtgcg agacccaggg tcggcgccgc tgcgtttccc
      721 ggaaccacgc ccagagcagc cgcgtccctg cgcaaaccca gggctgcctt ggaaaaggcg
      781 caaccccaac ccctttcaag acctagctag cgaattcaaa aaagcaccga ctcggtgcca
      841 ctttttcaag ttgataacgg actagcctta ttttaacttg ctatttctag ctctaaaacg
      901 ctgaagcact gcacgccatc ggtgtttcgt cctttccaca agatatataa agccaagaaa
      961 tcgaaatact ttcaagttac ggtaagcata tgatagtcca ttttaaaaca taattttaaa
     1021 actgcaaact acccaagaaa ttattacttt cctacgtcac gtattttgta ctaatatctt
     1081 tggtgtttac agtccaaatt aattctaaat tatctcccct acaagccctg gtatccgaaa
     1141 atgcaaatat gaaaggaatc atgggaaaat aggcccttcg gtaaccttaa ttaaaccaaa
     1201 actggaatcc ttggctgtcc cgtggaaaag aacccgaaaa attttgaaat tntgtgannt
     1261 tgcgtattcg gaattcctta aaatttgaaa ggtccggttg gcctaataag tccaacaaat
     1321 cctttccccc tgaanttgaa accccccaat cccacccntt tcaattttaa aaaattggng
     1381 aggaaaaaac gggccgtttt ttgccaggca aattaggcgg acgccagtnt ccgcaattcc
     1441 tnaatctgga acnggaaagg aatgtggang cctcaggttc cagtcccaac cacaatgtct
     1501 ctgtaagata accggaagng cgaccgnaga ccccgcctcc agaggtng
//
