LOCUS       sequence-68898-e         929 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Human pSico-based U6 vector containing murine U6 promoter and sgRNA 
            targeting endogenous CD71 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 929)
  AUTHORS   Gilbert LA, Larson MH, Morsut L, Liu Z, Brar GA, Torres SE, 
            Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA, 
            Weissman JS, Qi LS
  TITLE     CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in 
            Eukaryotes.
  JOURNAL   Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi: 
            10.1016/j.cell.2013.06.044.
  PUBMED    23849981
REFERENCE   2  (bases 1 to 929)
  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: "Cell. 2013 
            Jul 9. pii: S0092-8674(13)00826-X. doi: 10.1016/j.cell.2013.06.044."
COMMENT     Sequence Label: pU6-sgCD71-2
FEATURES             Location/Qualifiers
     source          1..929
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             58..111
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     regulatory      118..127
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             124..834
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(269..287)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     537..556
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(854..887)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc ccggtgccgg atcgggagag
       61 ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc accggtcgcc
      121 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
      181 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
      241 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
      301 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
      361 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
      421 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
      481 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
      541 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
      601 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
      661 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
      721 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
      781 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtaagaattc
      841 gtcgagggac ctaataactt cgtatagcat acattatacg aagttataca tgtttaaggg
      901 ttccggttcc actaggtaca attcgatat
//
