LOCUS       sequence-112391-        1090 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 1090)
  TITLE     Staphylococcus aureus Cas9
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1090)
  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            "Unpublished"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..1090
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     216..233
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    350..371
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(386..391,392..409,410..416)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    424..440
                     /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)."
     primer_bind     429..451
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     448..464
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     448..464
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    543..567
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(579..654)
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     promoter        complement(673..921)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(731..750)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(901..921)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(1060..1079)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1061..1079)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
       61 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      121 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      181 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      241 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      301 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
      361 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
      421 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
      481 ttggtaccga gctcggatcc actagtaacg gccgccagtg tgctggaatt cgccctttaa
      541 tgccaacttt gtacaagaaa gctgggtcta gaaaaaaatc tcgccaacaa gttgacgaga
      601 taaacacggc attttgcctt gttttagtag attctgtttc cagagtacta aaacaggtct
      661 tctcgaagac ccggtgtttc gtcctttcca caagatatat aaagccaaga aatcgaaata
      721 ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa
      781 ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac
      841 agtcaaatta attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg
      901 aatcatggga aataggccct cttcctgccc gaccttggta ccggatccag tcgactgaat
      961 tggttccttt aaagcctgct tttttgtaca aagggcgaat tctgcagata tccatcacac
     1021 tggcggccgc tcgagcatgc atctagaggg cccaattcgc cctatagtga gtcgtattac
     1081 aattcactgg
//
