LOCUS       sequence-146906-         849 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA1-boxB-MS2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 849)
  AUTHORS   Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T
  TITLE     Multiplexed labeling of genomic loci with dCas9 and engineered 
            sgRNAs using CRISPRainbow.
  JOURNAL   Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526.
  PUBMED    27088723
REFERENCE   2  (bases 1 to 849)
  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: "Nat 
            Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526."
COMMENT     Sequence Label: pLH-sgRNA1-boxB-MS2
FEATURES             Location/Qualifiers
     source          1..849
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             353..658
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     537..556
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    691..709
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     misc_RNA        751..769
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
ORIGIN
        1 ggacgaaaca ccggtgtctt cgaggcttac taaaagccag ataacagtat gcgtatttgc
       61 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
      121 gaggtgtgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
      181 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
      241 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
      301 cggtttattg aaatgaacgg ctcttttgct gacgagaaca gggactggtg aaatgcagtt
      361 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
      421 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
      481 agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
      541 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
      601 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaac
      661 cctgcaggac gaagacccgt ttgagagcta gggccctgaa gaagggccct agcaagttca
      721 aataaggcta gtccgttatc aacttggcca acatgaggat cacccatgtc tgcagggcca
      781 agtggcaccg agtcggtgct ttttttgaat tctcgacctc gagacaaatg gcagtattca
      841 tccacaatt
//
