LOCUS       sequence-135171-        1087 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses S. pyogenes CRISPR chimeric RNA element with customizable 
            sgRNA from U6 promoter and Crimson reporter from EF-1a promoter. 
            Lentiviral backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1087)
  AUTHORS   Canver MC, Smith EC, Sher F, Pinello L, Sanjana NE, Shalem O, Chen 
            DD, Schupp PG, Vinjamur DS, Garcia SP, Luc S, Kurita R, Nakamura Y, 
            Fujiwara Y, Maeda T, Yuan GC, Zhang F, Orkin SH, Bauer DE
  TITLE     BCL11A enhancer dissection by Cas9-mediated in situ saturating 
            mutagenesis.
  JOURNAL   Nature. 2015 Sep 16. doi: 10.1038/nature15521.
  PUBMED    26375006
REFERENCE   2  (bases 1 to 1087)
  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: "Nature. 
            2015 Sep 16. doi: 10.1038/nature15521."
COMMENT     Sequence Label: lentiGuide-Crimson
FEATURES             Location/Qualifiers
     source          1..1087
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      24..33
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             30..707
                     /codon_start=1
                     /product="far-red noncytotoxic tetrameric variant of DsRed 
                     fluorescent protein (Strack et al., 2009)"
                     /label=E2-Crimson
                     /note="mammalian codon-optimized"
                     /translation="MDSTENVIKPFMRFKVHMEGSVNGHEFEIEGVGEGKPYEGTQTAK
                     LQVTKGGPLPFAWDILSPQFFYGSKAYIKHPADIPDYLKQSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGTLIYHVKFIGVNFPSDGPVMQKKTLGWEPSTERNYPRDGVLKGENHM
                     ALKLKGGGHYLCEFKSIYMAKKPVKLPGYHYVDYKLDITSHNEDYTVVEQYERAEARHH
                     LFQ"
     primer_bind     428..447
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(776..796)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
ORIGIN
        1 catttcaggt gtcgtgacgt acggccacca tggatagcac tgagaacgtc atcaagccct
       61 tcatgcgctt caaggtgcac atggagggct ccgtgaacgg ccacgagttc gagatcgagg
      121 gcgtgggcga gggcaagccc tacgagggca cccagaccgc caagctgcaa gtgaccaagg
      181 gcggccccct gcccttcgcc tgggacatcc tgtcccccca gttcttctac ggctccaagg
      241 cgtacatcaa gcaccccgcc gacatccccg actacctcaa gcagtccttc cccgagggct
      301 tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt ggtgaccgtg acccaggact
      361 cctccctgca ggacggcacc ctcatctacc acgtgaagtt catcggcgtg aacttcccct
      421 ccgacggccc cgtaatgcag aagaagactc tgggctggga gccctccact gagcgcaact
      481 acccccgcga cggcgtgctg aagggcgaga accacatggc gctgaagctg aagggcggcg
      541 gccactacct gtgtgagttc aagtccatct acatggccaa gaagcccgtg aagctgcccg
      601 gctaccacta cgtggactac aagctcgaca tcacctccca caacgaggac tacaccgtgg
      661 tggagcagta cgagcgcgcc gaggcccgcc accacctgtt ccagtagacg cgttaagtcg
      721 acaatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg
      781 ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc
      841 gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt
      901 tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca
      961 ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc
     1021 ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc
     1081 tgttggg
//
