LOCUS       sequence-159019-         806 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 806)
  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 806)
  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..806
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     149..166
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_RNA        complement(353..428)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(447..695)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(505..524)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(675..695)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(759..778)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(760..778)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(789..806)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(789..805)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
       61 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cattaggcac
      121 cccaggcttt acccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggagagcgc
      181 ccaatacgca aggaaacagc tatgaccatg ttaatgcagc tggcacgaca ggtttcccga
      241 ctggaaagcg ggcagtgagc ggaaggcccg tgaggccagt cttgtgctcc aggtaccacg
      301 cgttgtacag gtctcggtcc taacttgcta tttctagctc taaaacaaaa aagcaccgac
      361 tcggtgccac tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc
      421 tctaaaacag gtcttctcga agacccggtg tttcgtcctt tccacaagat atataaagcc
      481 aagaaatcga aatactttca agttacggta agcatatgat agtccatttt aaaacataac
      541 tttaaaactg caaactaccc aagaaattat tactttctac gtcacgtatt ttgtactaat
      601 atctttgtgt ttacagtcaa attaattcca attatctctc taacagcctt gtatcgtata
      661 tgcaaatatg aaggaatcat gggaaatagg ccctccatgg gagaccggat ccgagctctg
      721 gacaagacac gtggccttga cggccttccg ccaattcgcc ctatagtgag tcgtattacg
      781 tcgcgctcac tggccgtcgt tttaca
//
