LOCUS       sequence-146907-         875 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sgRNA1-boxB-MS2-PP7.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 875)
  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 875)
  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-PP7
FEATURES             Location/Qualifiers
     source          1..875
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             354..659
                     /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     538..557
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    692..710
                     /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        752..770
                     /label=MS2 stem loop
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
ORIGIN
        1 aggacgaaac accggtgtct tcgaggctta ctaaaagcca gataacagta tgcgtatttg
       61 cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa
      121 agaggtgtgc tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc
      181 tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc
      241 ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc
      301 ccggtttatt gaaatgaacg gctcttttgc tgacgagaac agggactggt gaaatgcagt
      361 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
      421 atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
      481 cagataaagt ctcccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
      541 tgatgaccac cgatatggcc agtgtgccgg tctccgttat cggggaagaa gtggctgatc
      601 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
      661 ccctgcagga cgaagacccg tttgagagct agggccctga agaagggccc tagcaagttc
      721 aaataaggct agtccgttat caacttggcc aacatgagga tcacccatgt ctgcagggcc
      781 aagtggcacc gagtcggtgc ggagcagacg atatggcgtc gctccttttt ttgaattctc
      841 gacctcgaga caaatggcag tattcatcca caatt
//
