LOCUS       sequence-139639-        1091 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  multiplex gRNA plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1091)
  AUTHORS   Port F, Bullock SL
  TITLE     Augmenting CRISPR applications in Drosophila with tRNA-flanked 
            sgRNAs.
  JOURNAL   Nat Methods. 2016 Oct;13(10):852-4. doi: 10.1038/nmeth.3972. Epub 
            2016 Sep 5.
  PUBMED    27595403
REFERENCE   2  (bases 1 to 1091)
  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"; doi: 
            "10.1038/nmeth.3972"; journalName: "Nat Methods"; date: "2016-10"; 
            volume: "13"; issue: "10"; pages: "852-4"
COMMENT     Sequence Label: pCFD5
FEATURES             Location/Qualifiers
     source          1..1091
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    95..116
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(131..136,137..154,155..161)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    169..185
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     174..196
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     193..209
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     193..209
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     228..248
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        230..248
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_RNA        complement(559..634)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_RNA        complement(721..796)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      121 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      181 aacaatttca cacaggaaac agctatgacc atgattacgc caagcgcgca attaaccctc
      241 actaaaggga acaaaagctg gagctcctgc aggttgttgg ttggcacacc acaaatatac
      301 tgttgccgag cacaattgtc tagaatgcat acgcattaag cgaacattaa aaagatgtga
      361 aaacataact attatgtcta aataaacaca cgtcagatgt atgtacgtca acggaaaacc
      421 attgtctata tattacaatt actaaataca taccaaattg aatacatatt gatgaaaaat
      481 aataaatact ggcgaaagca aaaaaacgaa acatttttat tttattgaac aactctcagg
      541 ctccaggtag gcaaaaaagc accgactcgg tgccactttt tcaagttgat aacggactag
      601 ccttatttta acttgctatt tctagctcta aaacaggtct tctgcaccag ccgggaatcg
      661 aacccgggtc tgtaccgtgg cagggtacta ttctaccact agaccactgg tgctttgtta
      721 gcaccgactc ggtgccactt tttcaagttg ataacggact agccttattt taacttgcta
      781 tttctagctc taaaacgaag accctgcatc ggccgggaat cgaacccggg ccgcccgcgt
      841 ggcaggcgag cattctacca ctgaaccacc gatgcttgat gtctacacac actcaaagcc
      901 ccgacgttaa attgaaaata ggtctatata tacgaactga gtctggaaaa agaagttgag
      961 aattataaaa agtagtgagc actggcgccc tctctgcttg gcgagctaac cttttcgcct
     1021 cttggctgag taggtggcgt ttcattctac tctgtaaaat taatgtagaa ttgaaacact
     1081 atggtcaaaa a
//
