LOCUS       sequence-112390-        1094 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cloning and expression of Streptococcus pyogenes guide RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1094)
  AUTHORS   Ding Q, Regan SN, Xia Y, Oostrom LA, Cowan CA, Musunuru K
  TITLE     Enhanced efficiency of human pluripotent stem cell genome editing 
            through replacing TALENs with CRISPRs.
  JOURNAL   Cell Stem Cell. 2013 Apr 4;12(4):393-4. doi: 
            10.1016/j.stem.2013.03.006.
  PUBMED    23561441
REFERENCE   2  (bases 1 to 1094)
  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.stem.2013.03"; journalName: "Cell Stem Cell"; date: 
            "2013-04-4- 4"; volume: "12"; issue: "4"; pages: "393-4"
COMMENT     Sequence Label: pGuide
FEATURES             Location/Qualifiers
     source          1..1094
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     199..221
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     213..230
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     214..230
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     237..256
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        237..255
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        398..646
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     398..418
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     569..588
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        665..740
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(752..776)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(864..882)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(865..882)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(900..916)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(900..916)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(913..935)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    924..940
                     /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(948..954,955..972,973..978))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    993..1014
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
       61 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
      121 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
      181 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattgtaat
      241 acgactcact atagggcgaa ttgggccctc tagatgcatg ctcgagcggc cgccagtgtg
      301 atggatatct gcagaattcg cccttcattg tacaaaaaag caggctttaa aggaaccaat
      361 tcagtcgact ggatccggta ccaaggtcgg gcaggaagag ggcctatttc ccatgattcc
      421 ttcatatttg catatacgat acaaggctgt tagagagata attagaatta atttgactgt
      481 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
      541 tgcagtttta aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta
      601 tttcgatttc ttggctttat atatcttgtg gaaaggacga aacaccgggt cttcgagaag
      661 acctgtttta gagctagaaa tagcaagtta aaataaggct agtccgttat caacttgaaa
      721 aagtggcacc gagtcggtgc tttttttcta gacccagctt tcttgtacaa agttggcatt
      781 aatgaagggc gaattccagc acactggcgg ccgttactag tggatccgag ctcggtacca
      841 agcttgatgc atagcttgag tattctatag tgtcacctaa atagcttggc gtaatcatgg
      901 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
      961 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1021 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     1081 ggccaacgcg cggg
//
