LOCUS       sequence-178907-         815 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  NHEJ donor for fusing a p2A-mCherry reporter to mouse Actb. EGFP 
            driven by EF1a promoter and U6-driven sgRNAs targeting Actb. AAV 
            backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 815)
  AUTHORS   Yao X, Wang X, Hu X, Liu Z, Liu J, Zhou H, Shen X, Wei Y, Huang Z, 
            Ying W, Wang Y, Nie YH, Zhang CC, Li S, Cheng L, Wang Q, Wu Y, Huang
            P, Sun Q, Shi L, Yang H
  TITLE     Homology-mediated end joining-based targeted integration using 
            CRISPR/Cas9.
  JOURNAL   Cell Res. 2017 May 19. doi: 10.1038/cr.2017.76.
  PUBMED    28524166
REFERENCE   2  (bases 1 to 815)
  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. 
            2017 May 19. doi: 10.1038/cr.2017.76."
COMMENT     Sequence Label: AAV_Actb NHEJ donor_U6_sgRNA_EF1a_GFP_polyA
FEATURES             Location/Qualifiers
     source          1..815
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             25..81
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             82..792
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(227..245)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     495..514
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 agtccgccta gaagcacttg cggcgccact aacttctccc tgttgaaaca agcaggggat
       61 gtcgaagaga atcccgggcc aatggtgagc aagggcgagg aggataacat ggccatcatc
      121 aaggagttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag
      181 atcgagggcg agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg
      241 accaagggtg gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc
      301 tccaaggcct acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc
      361 gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc
      421 caggactcct ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac
      481 ttcccctccg acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag
      541 cggatgtacc ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag
      601 gacggcggcc actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag
      661 ctgcccggcg cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac
      721 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag
      781 ctgtacaagt aaagtccgcc tagaagcact tgcgg
//
