LOCUS       sequence-137379-         800 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA scaffold and synthetic U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 800)
  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 800)
  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_sU6
FEATURES             Location/Qualifiers
     source          1..800
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(294..314)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     misc_RNA        complement(405..480)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(568..586)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(569..586)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(604..620)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(604..620)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(617..639)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    628..644
                     /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(652..658,659..676,677..682))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    697..718
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 cgagcggccg ccagtgtgat ggatatctgc agaattcgcc cttgaagacc caaaccaaac
       61 aaggcttttc tcgcatgcat atataaagtc tcaaaacaca caattacttt caagttacgg
      121 taagtttcct tttgtgctgt tttttaaaat aataatttag tatttgtatc tcttatagaa
      181 atccaagcct atcatgtaaa atgtagctag tattaaaaag aacagattat ctgtctttta
      241 tcgcacatta agcctctata gttactagga aatattatat gcaaatatga aggaatcatg
      301 ggaaataggc cctctctaga gccatttgtc tgcagaattg gcgcacgcgc taaaaacgga
      361 ctagccttat tttaacttgc tatttctagc tctaaaacaa aaaagcaccg actcggtgcc
      421 actttttcaa gttgataacg gactagcctt attttaactt gctatttcta gctctaaaac
      481 aggtcttcaa gggcgaattc cagcacactg gcggccgtta ctagtggatc cgagctcggt
      541 accaagcttg atgcatagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc
      601 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
      661 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
      721 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
      781 aatcggccaa cgcgcgggga
//
