LOCUS       sequence-114010-        1095 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  in vitro trancription of sgRNA targeting the zebrafish th locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1095)
  AUTHORS   Li J, Zhang BB, Ren YG, Gu SY, Xiang YH, Huang C, Du JL
  TITLE     Intron targeting-mediated and endogenous gene integrity-maintaining 
            knockin in zebrafish using the CRISPR/Cas9 system.
  JOURNAL   Cell Res. 2015 Apr 7. doi: 10.1038/cr.2015.43.
  PUBMED    25849248
REFERENCE   2  (bases 1 to 1095)
  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: "Cell Res. 
            2015 Apr 7. doi: 10.1038/cr.2015.43."
COMMENT     Sequence Label: pT7-th-sgRNA
FEATURES             Location/Qualifiers
     source          1..1095
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(205..309)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     377..395
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(433..455)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     555..574
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     768..790
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     782..799
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     783..799
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_RNA        complement(845..920)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(938..957)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(939..957)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1010..1026)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1010..1026)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1023..1045)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1034..1050
                     /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(1058..1064,1065..1082,1083..1088))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
       61 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
      121 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
      181 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
      241 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
      301 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat
      361 gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga
      421 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
      481 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
      541 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga
      601 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
      661 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      721 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      781 ttgtaaaacg acggccagtg ccaaagaagc atgacggcaa gtggacgatt aagctttaaa
      841 aaaagcaccg actcggtgcc actttttcaa gttgataacg gactagcctt attttaactt
      901 gctatttcta gctctaaaac ctttatgagt gtaacaaccc tatagtgagt cgtattagaa
      961 ttcaatctcc agaggatcgc cgggaaccga ggacgagttc gtaatcatgg tcatagctgt
     1021 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1081 agtgtaaagc ctggg
//
