LOCUS       sequence-68893-e         891 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Human pSico-based U6 vector containing murine U6 promoter and sgRNA 
            targeting GFP (NT1).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 891)
  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 891)
  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-sgGFP-NT1
FEATURES             Location/Qualifiers
     source          1..891
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             60..113
                     /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      120..129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             126..836
                     /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(271..289)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     539..558
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(856..889)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc ggatcgggag
       61 agggcagagg aagtctgcta acatgcggtg acgtcgagga gaatcctggc ccaccggtcg
      121 ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
      181 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      241 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
      301 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
      361 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
      421 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
      481 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
      541 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
      601 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
      661 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
      721 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
      781 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaagaat
      841 tcgtcgaggg acctaataac ttcgtatagc atacattata cgaagttata c
//
