LOCUS       sequence-159017-         823 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  BsaI-based cloning of SpCas9 gRNA guide sequence, position 2/12/22 
            in the array.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 823)
  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 823)
  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-g2
FEATURES             Location/Qualifiers
     source          1..823
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(189..293)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     594..616
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     608..625
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     609..625
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     636..655
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        636..654
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     719..739
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
       61 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
      121 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
      181 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
      241 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
      301 ccccgaaaag tgccacctaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
      361 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
      421 caaaagaata gaccgagata gggttgagtg gccgctacag ggcgctccca ttcgccattc
      481 aggctgcgca actgttggga agggcgtttc ggtgcgggcc tcttcgctat tacgccagct
      541 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
      601 acgacgttgt aaaacgacgg ccagtgagcg cgacgtaata cgactcacta tagggcgaat
      661 tggcggaagg ccgtcaaggc cacgtgtctt gtccagagct cggatccggt ctcccatgga
      721 gggcctattt cccatgattc cttcatattt gcatatacga tacaaggctg ttagagagat
      781 aattggaatt aatttgactg taaacacaaa gatattagta caa
//
