LOCUS       sequence-122027-         961 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Empty vector for the expression U6 driven gRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 961)
  AUTHORS   Mulholland CB, Smets M, Schmidtmann E, Leidescher S, Markaki Y, 
            Hofweber M, Qin W, Manzo M, Kremmer E, Thanisch K, Bauer C, Rombaut 
            P, Herzog F, Leonhardt H, Bultmann S
  TITLE     A modular open platform for systematic functional studies under 
            physiological conditions.
  JOURNAL   Nucleic Acids Res. 2015 May 24. pii: gkv550.
  PUBMED    26007658
REFERENCE   2  (bases 1 to 961)
  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: "Nucleic 
            Acids Res. 2015 May 24. pii: gkv550."
COMMENT     Sequence Label: pEX-A-U6-gRNA
FEATURES             Location/Qualifiers
     source          1..961
                     /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"
     protein_bind    486..510
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(522..597)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(650..898)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(708..727)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(878..898)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
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 gaattcggat ccaagcttgc ggccgcaggt
      481 taatgccaac tttgtacaag aaagctgggt ctagaaaaaa agcaccgact cggtgccact
      541 ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct ctaaaacggg
      601 tcagctactc ctcgcatctc gcatgcatcg catcgaggag atccgatccg gtgtttcgtc
      661 ctttccacaa gatatataaa gccaagaaat cgaaatactt tcaagttacg gtaagcatat
      721 gatagtccat tttaaaacat aattttaaaa ctgcaaacta cccaagaaat tattactttc
      781 tacgtcacgt attttgtact aatatctttg tgtttacagt caaattaatt ctaattatct
      841 ctctaacagc cttgtatcgt atatgcaaat atgaaggaat catgggaaat aggccctctt
      901 cctgcccgac cttggtaccg gatccagtcg actgaattgg ttcctttaaa gcctgctttt
      961 t
//
