LOCUS       sequence-71683-e         895 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-one plasmid targeting tdTomato.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 895)
  AUTHORS   Hou Z, Zhang Y, Propson NE, Howden SE, Chu LF, Sontheimer EJ, 
            Thomson JA
  TITLE     Efficient genome engineering in human pluripotent stem cells using 
            Cas9 from Neisseria meningitidis.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Aug 12.
  PUBMED    23940360
REFERENCE   2  (bases 1 to 895)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Aug 12."
COMMENT     Sequence Label: 
            pSimpleII-U6-tracr-U6-crRNA(tdTomato)-NLS-NmCas9-HA-NLS(s)
FEATURES             Location/Qualifiers
     source          1..895
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(191..295)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        complement(390..638)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(448..467)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(618..638)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     676..694
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     promoter        complement(713..731)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        complement(773..865)
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
ORIGIN
        1 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
       61 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
      121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
      181 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
      241 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
      301 ttccccgaaa agtgccacct gacgtcgttt aaacaaaaaa acgaaatgag aaagggagct
      361 acaacatgtc ggcggggtgc ttcacgtacg gtgtttcgtc ctttccacaa gatatataaa
      421 gccaagaaat cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat
      481 aattttaaaa ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact
      541 aatatctttg tgtttacagt caaattaatt ctaattatct ctctaacagc cttgtatcgt
      601 atatgcaaat atgaaggaat catgggaaat aggccctctt cctgcccgac cttagatccg
      661 aatccatatc gatatgctag ttattgctca gcggtggcag cagccaactc agcctatagt
      721 gagtcgtatt aatttcgcgg gatgtttaaa cgcggccgca aaaaaaccga aataaacgat
      781 gccccttaaa gcagaagctt taaggggcag agcgttgcgg cacatctttt cagacggcct
      841 tattgtagca acggttctca tttcggtgtt tcgtcctttc cacaagatat ataaa
//
