LOCUS       sequence-159028-         812 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  BsaI-based cloning of SpCas9 gRNA guide sequence, position 5/15/25 
            in the array.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 812)
  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 812)
  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-g5
FEATURES             Location/Qualifiers
     source          1..812
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     142..159
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_RNA        complement(346..421)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(440..688)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(498..517)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(668..688)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(752..771)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(753..771)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(782..799)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(782..798)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
       61 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcattagg caccccaggc
      121 tttacccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggagag cgcccaatac
      181 gcaaggaaac agctatgacc atgttaatgc agctggcacg acaggtttcc cgactggaaa
      241 gcgggcagtg agcggaaggc ccgtgaggcc agtcttgtgc tccaggtacc acgcgttgta
      301 caggtctcct gcataacttg ctatttctag ctctaaaaca aaaaagcacc gactcggtgc
      361 cactttttca agttgataac ggactagcct tattttaact tgctatttct agctctaaaa
      421 caggtcttct cgaagacccg gtgtttcgtc ctttccacaa gatatataaa gccaagaaat
      481 cgaaatactt tcaagttacg gtaagcatat gatagtccat tttaaaacat aattttaaaa
      541 ctgcaaacta cccaagaaat tattactttc tacgtcacgt attttgtact aatatctttg
      601 tgtttacagt caaattaatt ccaattatct ctctaacagc cttgtatcgt atatgcaaat
      661 atgaaggaat catgggaaat aggccctcaa caggagaccg gatccgagct ctggacaaga
      721 cacgtggcct tgacggcctt ccgccaattc gccctatagt gagtcgtatt acgtcgcgct
      781 cactggccgt cgttttacaa cgtcgtgact gg
//
