LOCUS       sequence-110539-         774 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Vector for expression of Nm sgRNA1.1 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 774)
  AUTHORS   Ma H, Naseri A, Reyes-Gutierrez P, Wolfe SA, Zhang S, Pederson T
  TITLE     Multicolor CRISPR labeling of chromosomal loci in human cells.
  JOURNAL   Proc Natl Acad Sci U S A. 2015 Mar 10;112(10):3002-7. doi: 
            10.1073/pnas.1420024112. Epub 2015 Feb 23.
  PUBMED    25713381
REFERENCE   2  (bases 1 to 774)
  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: 
            "10.1073/pnas.1420024112"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2015-03-10- 10"; volume: "112"; issue: "10"; pages: "3002-7"
COMMENT     Sequence Label: pLH-nmsgRNA1.1
FEATURES             Location/Qualifiers
     source          1..774
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    108..225
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     misc_RNA        complement(275..367)
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     CDS             complement(416..721)
                     /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="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(518..537)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 agcagccgcg tccctgcgca aacccagggc tgccttggaa aaggcgcaac cccaaccccc
       61 tcgagccgcg gccaaagtgg atctctgctg tccctgtaat aaacccgaaa attttgaatt
      121 tttgtaattt gtttttgtaa ttctttagtt tgtatgtctg ttgctattat gtctactatt
      181 ctttcccctg cactgtaccc cccaatcccc ccttttcttt taaaattgtg gatgaatact
      241 gccatttgtc tcgaggtcga gaattcaaaa aaattaaacg atgcccctta aagcagaagc
      301 tttaaggggc agagcgttgc ggcacatctt ttcagacggc cttattgtag caacggttct
      361 catttcgttt ccgaaatgag aaagggagct acaactggtc ttcgtcctgc agggtttata
      421 ttccccagaa catcaggtta atggcgtttt tgatgtcatt ttcgcggtgg ctgagatcag
      481 ccacttcttc cccgataacg gagaccggca cactggccat atcggtggtc atcatgcgcc
      541 agctttcatc cccgatatgc accaccgggt aaagttcacg ggagacttta tctgacagca
      601 gacgtgcact ggccaggggg atcaccatcc gtcgcccggg cgtgtcaata atatcactct
      661 gtacatccac aaacagacga taacggctct ctcttttata ggtgtaaacc ttaaactgca
      721 tttcaccagt ccctgttctc gtcagcaaaa gagccgttca tttcaataaa ccgg
//
