LOCUS       sequence-72726-e         757 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian U6-driven sgRNA (NMm1) targeting GTCCCCTCCACCCCACAGTG.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 757)
  AUTHORS   Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM
  TITLE     Orthogonal Cas9 proteins for RNA-guided gene regulation and editing.
  JOURNAL   Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681.
  PUBMED    24076762
REFERENCE   2  (bases 1 to 757)
  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 
            Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681."
COMMENT     Sequence Label: M-NM-sgRNA
FEATURES             Location/Qualifiers
     source          1..757
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        57..157
                     /label=Nm gRNA scaffold
                     /note="guide RNA scaffold for the Neisseria meningitidis 
                     CRISPR/Cas9 system (Esvelt et al., 2013)"
     protein_bind    complement(169..193)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(268..287)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(269..287)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(294..311)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(294..310)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(303..325)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             448..750
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
                     SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     629..648
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 ttggctttat atatcttgtg gaaaggacga aacaccgtcc cctccacccc acagtggttg
       61 tagctccctt tctcgaaaga gaaccgttgc tacaataagg ccgtctgaaa agatgtgccg
      121 caacgctctg ccccttaaag cttctgcttt aacgggcttt ttttctagac ccagctttct
      181 tgtacaaagt tggcattaaa gggcgaattc tgcagatatc catcacactg gcggccgctc
      241 gagcatgcat ctagagggcc caattcgccc tatagtgagt cgtattacaa ttcactggcc
      301 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
      361 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
      421 caacagttgc gcagcctata cgtacggcag tttaaggttt acacctataa aagagagagc
      481 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgccggg gcgacggatg
      541 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg
      601 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg
      661 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac
      721 gccattaacc tgatgttctg gggaatataa atgtcag
//
