LOCUS       sequence-71684-e         854 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 854)
  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 854)
  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..854
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     294..313
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        378..626
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     378..398
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     549..568
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        626..718
                     /label=Nm tracrRNA
                     /note="trans-activating CRISPR RNA for the Neisseria 
                     meningitidis CRISPR/Cas9 system (Hou et al., 2013)"
     promoter        760..778
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(797..815)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 aaggcggagc cagtacacga catcactttc ccagtttacc ccgcgccacc ttctctaggc
       61 accggttcaa ttgccgaccc ctccccccaa cttctcgggg actgtgggcg atgtgcgctc
      121 tgcccactga cgggcaccgg agcctccgcg aagcagcgca aaacgcctaa ccctaagcag
      181 attcttcatg caattgtcgg tcaagccttg ccttgttgta gcttaaattt tgctcgcgca
      241 ctactcagcg acctccaaca cacaagcagg gagcagatac tggcttaact atgcggcatc
      301 agagcagatt gtactgagag tgcaccatag gggatcggga gatctcccga tccgtcgacg
      361 tcaaggtcgg gcaggaagag ggcctatttc ccatgattcc ttcatatttg catatacgat
      421 acaaggctgt tagagagata attagaatta atttgactgt aaacacaaag atattagtac
      481 aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt
      541 ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat
      601 atatcttgtg gaaaggacga aacaccgaaa tgagaaccgt tgctacaata aggccgtctg
      661 aaaagatgtg ccgcaacgct ctgcccctta aagcttctgc tttaaggggc atcgtttatt
      721 tcggtttttt tgcggccgcg tttaaacatc ccgcgaaatt aatacgactc actataggct
      781 gagttggctg ctgccaccgc tgagcaataa ctagcatatc gatatggatt cggatctaag
      841 gtcgggcagg aaga
//
