LOCUS       sequence-223217-        1685 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPRi negative control. Catalytically inactive Cas9 from S. 
            pyogenes with 2A-BlastR under the EF1a core promoter, and sgRNA1 
            against the BFP CDS.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1685)
  TITLE     Ferrer Lab Plasmids
REFERENCE   2  (bases 1 to 1685)
  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"
COMMENT     Sequence Label: Lenti-(BB)-EF1a-KRAB-dCas9-P2A-BlastR BFP-guide1
FEATURES             Location/Qualifiers
     source          1..1685
                     /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        264..475
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     regulatory      488..497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             524..709
                     /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"
ORIGIN
        1 ttgnagtttt aaattatgtt ttaaatggac tatcatatgc ttaccgtaac ttgaaagtat
       61 ttcgatttct tggctttata tatcttgtgg aaaggacgaa acaccgatgg cgtgcagtgc
      121 ttcagcgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt atcaacttga
      181 aaaagtggca ccgagtcggt gcttttttga attcgctagc taggtcttga aaggagtggg
      241 aattggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt
      301 ggggggaggg gtcggcaatt gatccggtgc ctagagaagg tggcgcgggg taaactggga
      361 aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac cgtatataag
      421 tgcagtagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa cacaggaccg
      481 gtgatccgcc accatggatg ctaagtcact aactgcctgg tcccggacac tggtgacctt
      541 caaggatgta tttgtggact tcaccaggga ggagtggaag ctgctggaca ctgctcagca
      601 gatcgtgtac agaaatgtga tgctggagaa ctataagaac ctggtttcct tgggttatca
      661 gcttactaag ccagatgtga tcctccggtt ggagaaggga gaagagcccg gcggttccgg
      721 cggagggtcg atggacaaga agtacagcat cggcctggcc atcggcacca actctgtggg
      781 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
      841 caccgaccgg cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggaga
      901 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
      961 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
     1021 cttccacaga ctggaagagt ccttcctggt ggaagaagga taagaagcac gaaccggcac
     1081 cccatctttc ggcaacatcc tgggacgaag gtggcctacc acgagaaagt acccccacca
     1141 tctaccacct ggaaaaagga aactgggtgg acagcacccg aacaaggccg aacctgcggc
     1201 tgaatctatc cgggccctgg ccccacagga tcaagtttcc cggggccact tccnggaatc
     1261 agagggccaa ccttgacccc cggaaaaaag ncgaacttgg aagaaagctg gttactccag
     1321 gttgggggcc agacctaacc aaccagttgg ttccaagaga aaacgccatg caancgcacg
     1381 gacgtgggac aacccaaggc atcntccggt tctggcccaa taagnaaaaa aggaaaacaa
     1441 gtgtggagga aatacaaaat caccccaccc ttcgcgccgg gaaaaagana aaatagattc
     1501 gngttnccgg acactctgaa ggtgtccncg ganntcgggg gtgtcccgaa cccanatttn
     1561 anagganaag attttcgaga ctcggggcca aaaaggggga acttggnggt nntggaaaaa
     1621 gcgnnaacca aaaaggaaaa accgnanaag ngaaccggtg gngtatcaaa gagaagaaaa
     1681 acaag
//
