LOCUS       Exported                 669 bp ds-DNA     linear   SYN 17-MAY-2021
DEFINITION  Expresses the S. pyogenes sgRNA from the mouse U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pmU6-gRNA
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 669)
  AUTHORS   Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA
  TITLE     Multiplex CRISPR/Cas9-based genome engineering from a single 
            lentiviral vector.
  JOURNAL   Nucleic Acids Res. 2014 Aug 13. pii: gku749.
  PUBMED    25122746
REFERENCE   2  (bases 1 to 669)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 17, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..669
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_RNA        complement(55..130)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(148..461)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for mouse U6 snRNA (Das 
                     et al., 1988)"
     primer_bind     complement(209..229)
                     /label=mU6-F
                     /note="Mouse U6 promoter, forward primer"
     primer_bind     complement(486..505)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(487..505)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(515..532)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(515..531)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(524..546)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 tcactaaagg gaacaaaagc tggagctcca ccgcggcgtc tcagtacaaa aaaagcaccg
       61 actcggtgcc actttttcaa gttgataacg gactagcctt attttaactt gctatttcta
      121 gctctaaaac aggtcttctc gaagacaaac aaggcttttc tccaagggat atttatagtc
      181 tcaaaacaca caattacttt acagttaggg tgagtttcct tttgtgctgt tttttaaaat
      241 aataatttag tatttgtatc tcttatagaa atccaagcct atcatgtaaa atgtagctag
      301 tattaaaaag aacagattat ctgtctttta tcgcacatta agcctctata gttactagga
      361 aatattatat gcaaattaac cggggcaggg gagtagccga gcttctccca caagtctgtg
      421 cgagggggcc ggcgcgggcc tagagatggc ggcgtcggat ctatctgaga cgggtaccca
      481 attcgcccta tagtgagtcg tattacgcgc gctcactggc cgtcgtttta caacgtcgtg
      541 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      601 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      661 atggcgaat
//
