LOCUS       sequence-107939-         988 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA cloning backbone with MS2 loops at tetraloop and stemloop 2. 
            Contains BbsI sites for insertion of spacer sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 988)
  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 988)
  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: "Nature. 
            2014 Dec 10. doi: 10.1038/nature14136."
COMMENT     Sequence Label: sgRNA(MS2) cloning backbone
FEATURES             Location/Qualifiers
     source          1..988
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     131..150
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        244..262
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        314..332
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     rep_origin      444..899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(531..550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     741..762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(916..935)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
       61 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
      121 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
      181 atatatcttg tggaaaggac gaaacaccgg gtcttcgaga agacctgttt tagagctagg
      241 ccaacatgag gatcacccat gtctgcaggg cctagcaagt taaaataagg ctagtccgtt
      301 atcaacttgg ccaacatgag gatcacccat gtctgcaggg ccaagtggca ccgagtcggt
      361 gctttttttg aattctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
      421 gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
      481 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc
      541 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
      601 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
      661 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
      721 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat
      781 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa
      841 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt
      901 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
      961 gccaacaccc gctgacgcgc cctgacgg
//
