LOCUS       sequence-112392-        1046 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cloning and expression of Staphylococcus aureus guide RNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1046)
  TITLE     Staphylococcus aureus Cas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1046)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pSaGuide
FEATURES             Location/Qualifiers
     source          1..1046
                     /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        395..643
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     395..415
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     566..585
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        662..737
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     protein_bind    complement(749..773)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(852..868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(852..868)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(865..887)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    876..892
                     /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(900..906,907..924,925..930))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    945..966
                     /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 ccctttgtac aaaaaagcag gctttaaagg aaccaattca
      361 gtcgactgga tccggtacca aggtcgggca ggaagagggc ctatttccca tgattccttc
      421 atatttgcat atacgataca aggctgttag agagataatt agaattaatt tgactgtaaa
      481 cacaaagata ttagtacaaa atacgtgacg tagaaagtaa taatttcttg ggtagtttgc
      541 agttttaaaa ttatgtttta aaatggacta tcatatgctt accgtaactt gaaagtattt
      601 cgatttcttg gctttatata tcttgtggaa aggacgaaac accgggtctt cgagaagacc
      661 tgttttagta ctctggaaac agaatctact aaaacaaggc aaaatgccgt gtttatctcg
      721 tcaacttgtt ggcgagattt ttttctagac ccagctttct tgtacaaagt tggcattaaa
      781 gggcgaattc cagcacactg gcggccgtta ctagtggatc cgagctcggt accaagcttg
      841 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
      901 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
      961 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     1021 cattaatgaa tcggccaacg cgcggg
//
