LOCUS       sequence-152736-         973 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes an sgRNA for spCas9 driven by hU6 promoter with a modified 
            scaffold (Chen et al. Cell 2013) and BbsI sites that allow insertion
            of a spacer sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 973)
  AUTHORS   Nelles DA, Fang MY, O'Connell MR, Xu JL, Markmiller SJ, Doudna JA, 
            Yeo GW
  TITLE     Programmable RNA Tracking in Live Cells with CRISPR/Cas9.
  JOURNAL   Cell. 2016 Mar 16. pii: S0092-8674(16)30204-5. doi: 
            10.1016/j.cell.2016.02.054.
  PUBMED    26997482
REFERENCE   2  (bases 1 to 973)
  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: "Cell. 2016 
            Mar 16. pii: S0092-8674(16)30204-5. doi: 
            10.1016/j.cell.2016.02.054."
COMMENT     Sequence Label: pBluescriptSKII+ U6-sgRNA(F+E) empty
FEATURES             Location/Qualifiers
     source          1..973
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     rep_origin      complement(50..505)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(187..208)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     399..418
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     632..654
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     646..663
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     647..663
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     673..692
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        673..691
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     716..732
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     717..733
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     818..838
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc
       61 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
      121 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
      181 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
      241 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
      301 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
      361 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
      421 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
      481 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa ctgttgggaa
      541 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
      601 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
      661 agtgagcgcg cgtaatacga ctcactatag ggcgaattgg gtaccgggcc ccccctcgag
      721 gtcgacggta tcgataagct tgatatcgtg tacaaaaaag caggctttaa aggaaccaat
      781 tcagtcgact ggatccggta ccaaggtcgg gcaggaagag ggcctatttc ccatgattcc
      841 ttcatatttg catatacgat acaaggctgt tagagagata attagaatta atttgactgt
      901 aaacacaaag atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt
      961 tgcagtttta aaa
//
