LOCUS       sequence-82775-e         794 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 794)
  AUTHORS   Jao LE, Wente SR, Chen W
  TITLE     Efficient multiplex biallelic zebrafish genome editing using a 
            CRISPR nuclease system.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Aug 5.
  PUBMED    23918387
REFERENCE   2  (bases 1 to 794)
  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: "Proc Natl 
            Acad Sci U S A. 2013 Aug 5."
COMMENT     Sequence Label: pT3TS-nCas9n
FEATURES             Location/Qualifiers
     source          1..794
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     186..203
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    320..341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(356..361,362..379,380..386)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    394..410
                     /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     399..421
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     418..434
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     418..434
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        455..473
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     5'UTR           498..540
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      556..565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             574..594
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
       61 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
      121 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
      181 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
      241 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
      301 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
      361 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
      421 gaaacagcta tgaccatgat tacgccaagc tcgaaattaa ccctcactaa agggaacaaa
      481 agctggagct aagcttgctt gttctttttg cagaagctca gaataaacgc tcaactttgg
      541 cagatctaac tcgacgccac catggcttct ccacctaaga agaagagaaa ggtgggaagc
      601 atggataaga agtatagcat cggcctggat attggaacta actccgtggg ttgggcagtg
      661 attacagacg actacaaggt ccctagcaag aaatttaagg tgctgggtaa caccgacagg
      721 cacagcatca agaaaaatct gattggagcc ctgctgttcg gttctggaga gactgccgaa
      781 gcaacacgcc tgaa
//
