LOCUS       sequence-150780-         996 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-one CRISPR/Cas9 vector with high-fidelity eSpCas9 expression 
            in human pluripotent stem cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 996)
  TITLE     pCAG-SpCas9-U6-gRNA
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 996)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pCAG-eCas9-GFP-U6-gRNA
FEATURES             Location/Qualifiers
     source          1..996
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        124..372
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     124..144
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     295..314
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        391..466
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     enhancer        646..949
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
       61 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      121 tgtgagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga
      181 gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt
      241 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat
      301 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttgtggaaa
      361 ggacgaaaca ccgggtcttc gagaagacct gttttagagc tagaaatagc aagttaaaat
      421 aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt ttgttttaga
      481 gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc caattctgca
      541 gacaaatggc tctagtaacg gccgccagtg tgctggaatt caggctagtt attaatagta
      601 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      661 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      721 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      781 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      841 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      901 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
      961 ccccacgttc tgcttcactc tccccatctc cccccc
//
