LOCUS       sequence-137374-         840 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA scaffold and mouse U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 840)
  AUTHORS   Vidigal JA, Ventura A
  TITLE     Rapid and efficient one-step generation of paired gRNA CRISPR-Cas9 
            libraries.
  JOURNAL   Nat Commun. 2015 Aug 17;6:8083. doi: 10.1038/ncomms9083.
  PUBMED    26278926
REFERENCE   2  (bases 1 to 840)
  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: "Nat 
            Commun."; date: "2015-08-17"; volume: "6:8083. doi"; pages: " 
            10.1038/ncomms9083"
COMMENT     Sequence Label: pDonor_mU6
FEATURES             Location/Qualifiers
     source          1..840
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(57..370)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     complement(118..138)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     misc_RNA        complement(467..542)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(630..648)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(631..648)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(666..682)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(666..682)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(679..701)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    690..706
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(714..720,721..738,739..744))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    759..780
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gagcggccgc cagtgtgatg gatatctgca gaattcgccc ttgaagaccc aaaccaaaca
       61 aggcttttct cgcatgcata tttatagtct caaaacacac aattacttta cagttagggt
      121 gagtttcctt ttgtgctgtt ttttaaaata ataatttagt atttgtatct cttatagaaa
      181 tccaagccta tcatgtaaaa tgtagctagt attaaaaaga acagattatc tgtcttttat
      241 cgcacattaa gcctctatag ttactaggaa atattatatg caaattaacc ggggcagggg
      301 agtagccgag cttctcccac aagtctgtgc gagggggccg gcgcgggcct agagatggcg
      361 gcgtcggatc tctagatcta gagccatttg tctgcagaat tggcgcacgc gctaaaaacg
      421 gactagcctt attttaactt gctatttcta gctctaaaac aaaaaagcac cgactcggtg
      481 ccactttttc aagttgataa cggactagcc ttattttaac ttgctatttc tagctctaaa
      541 acaggtcttc aagggcgaat tccagcacac tggcggccgt tactagtgga tccgagctcg
      601 gtaccaagct tgatgcatag cttgagtatt ctatagtgtc acctaaatag cttggcgtaa
      661 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
      721 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
      781 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
//
