LOCUS       sequence-159015-         801 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  BsaI-based cloning of SpCas9 gRNA guide sequence, position 1/11/21 
            in the array.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 801)
  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 801)
  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-g1
FEATURES             Location/Qualifiers
     source          1..801
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     148..165
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_RNA        complement(352..427)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(446..694)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(504..523)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(674..694)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(758..777)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(759..777)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
       61 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc attaggcacc
      121 ccaggcttta cccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggagagcgcc
      181 caatacgcaa ggaaacagct atgaccatgt taatgcagct ggcacgacag gtttcccgac
      241 tggaaagcgg gcagtgagcg gaaggcccgt gaggccagtc ttgtgctcca ggtaccacgc
      301 gttgtacagg tctcccatgt aacttgctat ttctagctct aaaacaaaaa agcaccgact
      361 cggtgccact ttttcaagtt gataacggac tagccttatt ttaacttgct atttctagct
      421 ctaaaacagg tcttctcgaa gacccggtgt ttcgtccttt ccacaagata tataaagcca
      481 agaaatcgaa atactttcaa gttacggtaa gcatatgata gtccatttta aaacataatt
      541 ttaaaactgc aaactaccca agaaattatt actttctacg tcacgtattt tgtactaata
      601 tctttgtgtt tacagtcaaa ttaattccaa ttatctctct aacagccttg tatcgtatat
      661 gcaaatatga aggaatcatg ggaaataggc cctcatagcg agaccggatc cgagctctgg
      721 acaagacacg tggccttgac ggccttccgc caattcgccc tatagtgagt cgtattacgt
      781 cgcgctcact ggccgtcgtt t
//
