LOCUS       sequence-159045-         799 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  BsaI-based cloning of SpCas9 gRNA guide sequence, position 10/20/30 
            in the array.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 799)
  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 799)
  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-g10
FEATURES             Location/Qualifiers
     source          1..799
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     146..163
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_RNA        complement(350..425)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(444..692)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(502..521)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(672..692)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(756..775)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(757..775)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
       61 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcat taggcacccc
      121 aggctttacc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg agagcgccca
      181 atacgcaagg aaacagctat gaccatgtta atgcagctgg cacgacaggt ttcccgactg
      241 gaaagcgggc agtgagcgga aggcccgtga ggccagtctt gtgctccagg taccacgcgt
      301 tgtacaggtc tcccgcctaa cttgctattt ctagctctaa aacaaaaaag caccgactcg
      361 gtgccacttt ttcaagttga taacggacta gccttatttt aacttgctat ttctagctct
      421 aaaacaggtc ttctcgaaga cccggtgttt cgtcctttcc acaagatata taaagccaag
      481 aaatcgaaat actttcaagt tacggtaagc atatgatagt ccattttaaa acataatttt
      541 aaaactgcaa actacccaag aaattattac tttctacgtc acgtattttg tactaatatc
      601 tttgtgttta cagtcaaatt aattccaatt atctctctaa cagccttgta tcgtatatgc
      661 aaatatgaag gaatcatggg aaataggccc tcgagcggag accggatccg agctctggac
      721 aagacacgtg gccttgacgg ccttccgcca attcgcccta tagtgagtcg tattacgtcg
      781 cgctcactgg ccgtcgttt
//
