LOCUS       sequence-126526-        1013 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of customizable guide RNA (gRNA) under cotnrol of murine 
            U6 promoter, also contains a CMV-puro-t2A-mCherry expression 
            cassette for mammalian gene knockdown.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1013)
  AUTHORS   Qi LS, Larson MH, Gilbert LA, Doudna JA, Weissman JS, Arkin AP, Lim 
            WA
  TITLE     Repurposing CRISPR as an RNA-Guided Platform for Sequence-Specific 
            Control of Gene Expression.
  JOURNAL   Cell. 2013 Feb 28;152(5):1173-83. doi: 10.1016/j.cell.2013.02.022.
  PUBMED    23452860
REFERENCE   2  (bases 1 to 1013)
  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"; doi: 
            "10.1016/j.cell.2013.02"; journalName: "Cell"; date: "2013-02-28- 
            28"; volume: "152"; issue: "5"; pages: "1173-83"
COMMENT     Sequence Label: pgRNA-humanized
FEATURES             Location/Qualifiers
     source          1..1013
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(191..224)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     228..248
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        331..406
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        531..834
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     989..1009
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 cgacgcgcca tctctaggcc cgcgccggcc ccctcgcacg gacttgtggg agaagctcgg
       61 ctactcccct gccccggtta atttgcatat aatatttcct agtaactata gaggcttaat
      121 gtgcgataaa agacagataa tctgttcttt ttaatactag ctacatttta catgataggc
      181 ttggatttct ataacttcgt atagcataca ttatacgaag ttataaacag cacaaaagga
      241 aactcaccct aactgtaaag taattgtgtg ttttgagact ataagtatcc cttggagaac
      301 caccttgttg gaccaggatg ggcaccaccc gttttagagc tagaaatagc aagttaaaat
      361 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttctcgagt
      421 actaggatcc attaggcggc cgcgtggata accgtattac cgccatgcat tagttattaa
      481 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
      541 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      601 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      661 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      721 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      781 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      841 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      901 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      961 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acg
//
