LOCUS       sequence-159039-         734 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  BsaI-based cloning of SpCas9 gRNA guide sequence, position 8/18/28 
            in the array.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 734)
  AUTHORS   Vad-Nielsen J, Lin L, Bolund L, Nielsen AL, Luo Y
  TITLE     Golden Gate Assembly of CRISPR gRNA expression array for 
            simultaneously targeting multiple genes.
  JOURNAL   Cell Mol Life Sci. 2016 May 13.
  PUBMED    27178736
REFERENCE   2  (bases 1 to 734)
  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 Mol 
            Life Sci. 2016 May 13."
COMMENT     Sequence Label: pMA-SpCas9-g8
FEATURES             Location/Qualifiers
     source          1..734
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     145..162
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_RNA        complement(349..424)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(443..691)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(501..520)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(671..691)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
       61 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcatt aggcacccca
      121 ggctttaccc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga gagcgcccaa
      181 tacgcaagga aacagctatg accatgttaa tgcagctggc acgacaggtt tcccgactgg
      241 aaagcgggca gtgagcggaa ggcccgtgag gccagtcttg tgctccaggt accacgcgtt
      301 gtacaggtct cttcgataac ttgctatttc tagctctaaa acaaaaaagc accgactcgg
      361 tgccactttt tcaagttgat aacggactag ccttatttta acttgctatt tctagctcta
      421 aaacaggtct tctcgaagac ccggtgtttc gtcctttcca caagatatat aaagccaaga
      481 aatcgaaata ctttcaagtt acggtaagca tatgatagtc cattttaaaa cataatttta
      541 aaactgcaaa ctacccaaga aattattact ttctacgtca cgtattttgt actaatatct
      601 ttgtgtttac agtcaaatta attccaatta tctctctaac agccttgtat cgtatatgca
      661 aatatgaagg aatcatggga aataggccct ctttcggaga ccggatccga gctctggaca
      721 agacacgtgg cctt
//
