LOCUS       Exported                 932 bp ds-DNA     linear   SYN 18-MAY-2021
DEFINITION  sgRNA cloning backbone with MS2 loops at tetraloop and stemloop 2. 
            Contains BbsI sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    sgRNA(MS2) cloning backbone
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 932)
  AUTHORS   Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena 
            C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F
  TITLE     Genome-scale transcriptional activation by an engineered CRISPR-Cas9
            complex.
  JOURNAL   Nature. 2014 Dec 10. doi: 10.1038/nature14136.
  PUBMED    25494202
REFERENCE   2  (bases 1 to 932)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 18, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..932
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_RNA        complement(91..109)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        complement(161..179)
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     promoter        complement(223..463)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     complement(273..292)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(443..463)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(606..625)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagaa ttcaaaaaaa
       61 gcaccgactc ggtgccactt ggccctgcag acatgggtga tcctcatgtt ggccaagttg
      121 ataacggact agccttattt taacttgcta ggccctgcag acatgggtga tcctcatgtt
      181 ggcctagctc taaaacaggt cttctcgaag acccggtgtt tcgtcctttc cacaagatat
      241 ataaagccaa gaaatcgaaa tactttcaag ttacggtaag catatgatag tccattttaa
      301 aacataattt taaaactgca aactacccaa gaaattatta ctttctacgt cacgtatttt
      361 gtactaatat ctttgtgttt acagtcaaat taattccaat tatctctcta acagccttgt
      421 atcgtatatg caaatatgaa ggaatcatgg gaaataggcc ctcgctagct gagcaaaagg
      481 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      541 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      601 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      661 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      721 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      781 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      841 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      901 agcgaggtat gtaggcggtg ctacagagtt ct
//
