LOCUS       sequence-178910-         879 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  MMEJ 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 879)
  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 879)
  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 MMEJ donor_U6_sgRNA_EF1a_GFP_polyA
FEATURES             Location/Qualifiers
     source          1..879
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             61..117
                     /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             118..828
                     /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(263..281)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     531..550
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 agtccgccta gaagcacttg cggcccctcc atcgtgcacc gcaagtgctt cttaattaac
       61 gccactaact tctccctgtt gaaacaagca ggggatgtcg aagagaatcc cgggccaatg
      121 gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg cttcaaggtg
      181 cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg cgagggccgc
      241 ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc cctgcccttc
      301 gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt gaagcacccc
      361 gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg ggagcgcgtg
      421 atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct gcaggacggc
      481 gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg ccccgtaatg
      541 cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga ggacggcgcc
      601 ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta cgacgctgag
      661 gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta caacgtcaac
      721 atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca gtacgaacgc
      781 gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagg cgcgccgcgg
      841 actgttactg agctgcccgc aagtgcttct aggcggact
//
