LOCUS       sequence-79289-e         946 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  gRNA expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 946)
  AUTHORS   Guo X, Zhang T, Hu Z, Zhang Y, Shi Z, Wang Q, Cui Y, Wang F, Zhao H,
            Chen Y
  TITLE     Efficient RNA/Cas9-mediated genome editing in Xenopus tropicalis.
  JOURNAL   Development. 2014 Jan 8.
  PUBMED    24401372
REFERENCE   2  (bases 1 to 946)
  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: 
            "Development. 2014 Jan 8."
COMMENT     Sequence Label: pUC57-Simple-gRNA backbone
FEATURES             Location/Qualifiers
     source          1..946
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_RNA        complement(43..118)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(137..155)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(220..236)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(220..236)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(233..255)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    244..260
                     /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)."
     promoter        complement(join(268..274,275..292,293..298))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    313..334
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(451..468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(702..721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcggtactt cgcgaatgcg tcgagatatt gggtctttaa aagcaccgac tcggtgccac
       61 tttttcaagt tgataacgga ctagccttat tttaacttgc tatttctagc tctaaaacag
      121 gtcttctcga agacgaccta tagtgagtcg tattaattgg gtatcggatg ccgggaccga
      181 cgagtgcaga ggcgtgcaag cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
      241 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
      301 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
      361 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
      421 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      481 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      541 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      601 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      661 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
      721 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
      781 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
      841 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
      901 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggta
//
