LOCUS       sequence-156779-        1030 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  U6 promoter driven sgRNA expression plasmid. Annealed 
            oligonucleotides encoding crRNA sequence are ligated into BbsI 
            site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1030)
  AUTHORS   Howden SE, McColl B, Glaser A, Vadolas J, Petrou S, Little MH, 
            Elefanty AG, Stanley EG
  TITLE     A Cas9 Variant for Efficient Generation of Indel-Free Knockin or 
            Gene-Corrected Human Pluripotent Stem Cells.
  JOURNAL   Stem Cell Reports. 2016 Sep 13;7(3):508-17. doi: 
            10.1016/j.stemcr.2016.07.001. Epub 2016 Aug 4.
  PUBMED    27499201
REFERENCE   2  (bases 1 to 1030)
  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: 
            "10.1016/j.stemcr.2016.07.001"; journalName: "Stem Cell Reports"; 
            date: "2016-09-13- 13"; volume: "7"; issue: "3"; pages: "508-17"
COMMENT     Sequence Label: pSMART-sgRNA (Sp)
FEATURES             Location/Qualifiers
     source          1..1030
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(47..149)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(150..181)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     misc_RNA        complement(274..349)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(375..623)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(433..452)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(603..623)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     terminator      732..761
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
ORIGIN
        1 ttaatcgcgg cctcgagcaa gacgtttccc gttgaatatg gctcatttta gcttccttag
       61 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
      121 gaaagttgga acctcttacg tgccgatcaa gtcaaaagcc tccggtcgga ggcttttgac
      181 tttctgctat ggaggtcagg tatgatttaa atggtcagta ttgagcgata tctagagaat
      241 tcgtccgaat ttcaagtgcg gccgcaaaaa aaagcaccga ctcggtgcca ctttttcaag
      301 ttgataacgg actagcctta ttttaacttg ctatttctag ctctaaaact tgtcttctgg
      361 aataatgaag acccggtgtt tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa
      421 tactttcaag ttacggtaag catatgatag tccattttaa aacataattt taaaactgca
      481 aactacccaa gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt
      541 acagtcaaat taattctaat tatctctcta acagccttgt atcgtatatg caaatatgaa
      601 ggaatcatgg gaaataggcc ctcttcctgc ccgaccttga cgtcgatatc ggatccgagc
      661 ttggattcaa gcttgatatc attcaggacg agcctcagac tccagcgtaa ctggactgca
      721 atcaactcac tggctcacct tcacgggtgg gcctttcttc ggtagaaaat caaaggatct
      781 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      841 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgag gtaactggct
      901 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
      961 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     1021 ctgccagtgg
//
