LOCUS       sequence-114009-        1097 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 1097)
  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 1097)
  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..1097
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     214..231
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    348..369
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(384..389,390..407,408..414)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    422..438
                     /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     427..449
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     446..462
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     446..462
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     515..534
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        515..533
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        552..627
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(673..690)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(673..689)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(682..704)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(898..917)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1017..1039
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1077..1095)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
       61 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      121 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      181 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      241 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
      301 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      361 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      421 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgaactcgtc
      481 ctcggttccc ggcgatcctc tggagattga attctaatac gactcactat agggttgtta
      541 cactcataaa ggttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
      601 cttgaaaaag tggcaccgag tcggtgcttt ttttaaagct taatcgtcca cttgccgtca
      661 tgcttctttg gcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac
      721 ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc
      781 ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg
      841 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
      901 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
      961 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     1021 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     1081 cgtgatacgc ctatttt
//
