LOCUS       sequence-178931-        2437 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  HMEJ donor for fusing a p2A-mCherry reporter to mouse Nanog. AAV 
            backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2437)
  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 2437)
  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_Nanog HMEJ donor
FEATURES             Location/Qualifiers
     source          1..2437
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             839..895
                     /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             896..1606
                     /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(1041..1059)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1309..1328
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 cgtaagtctc atatttcacc tggccccctc catcttacca tcttatttta tgtgtctgtg
       61 ccgtgtgtgt gtgtgtgtgt gtgtgtgtgg ctgacctatg gctgttgggt atggatccca
      121 gatttcctgt aagagcagtc agtggctctg cctttgcttc ccagtgtcat cctgatccat
      181 ctatgtccat tagtcctgtc cctttcccat ccctcttcct ccacgcactc tttcttgtga
      241 tctttggttc tttgaggcag gctctctcta tgtagccctg gcagtcagag atctgcctgc
      301 ctagtctcag gagtgctggg gttaacggcc tgtgcggcca tgctaatctt ttcagtattt
      361 ctgcagttta catctttaga gtcccctttt cactcattgc attaatcacc tctttcagaa
      421 gggctcagca ccagtggagt atcccagcat ccattgcagc tatccccagg gctatctggt
      481 gaacgcatct ggaagccttt ccatgtgggg cagccagact tggaccaacc caacttggag
      541 cagccagacc tggaccaacc caacttggaa caaccagacc tggaccaacc caacttggag
      601 cagccaggcc tggaccgctc agtcctggaa cggccagcct tggaatgctg ctccgctcca
      661 taacttcggg gaggactttc tgcagcctta cgtacagttg cagcaaaact tctctgccag
      721 tgatttggag gtgaatttgg aagccactag ggaaagccat gcgcatttta gcaccccaca
      781 agccttggaa ttattcctga actactctgt gactccacca ggtgaaatat taattaacgc
      841 cactaacttc tccctgttga aacaagcagg ggatgtcgaa gagaatcccg ggccaatggt
      901 gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct tcaaggtgca
      961 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
     1021 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc
     1081 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
     1141 cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat
     1201 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
     1261 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
     1321 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
     1381 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgctgaggt
     1441 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca acgtcaacat
     1501 caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgaacgcgc
     1561 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtaaggcg cgccgaaata
     1621 tgagacttac gcaacatctg ggcttaaagt cagggcaaag ccaggttcct tccttcttcc
     1681 aaatattttc atattttttt taaagattta tttattcatt atatgtaagt acactgtagc
     1741 tgtcttcaga cactccagaa gagggcgtca gatcttgtta cgtatggttg tgagccacca
     1801 tgtggttgct gggatttgaa ctcctgacct tcggaagagc agtcgggtgc tcttatccac
     1861 tgagccatct caccagcccc tggtttattt ttttaattat tatttgcttt ttgtttatca
     1921 agacagggtt tctctgcata gctctaattg tctttgaact agctctgcag accagcctgg
     1981 ccttgaactc agagatctgc ccacttatct ttgcctcctg aatgctggga ccaaaggtgg
     2041 cataccacca cacctggcat atatattgtt tatttctatt tctattttta ttggtgccag
     2101 agcaaaccta ggacttagaa catgctgggc accaactcaa cttctgagct ctatttacaa
     2161 cttggtgtgt tagtgtattt gtcttagttc tgaatttgtc ctttttttag tgttaactct
     2221 aggctttgga gacagtgagg tgcatatact ctctccttcc caagaataag tgcttgaaca
     2281 cccttaccca cgcccaccca cccatgctag tcttttttct tagaagcgtg ggtcttggta
     2341 tacactgtgt cattttgagg ggtgaggttt aaaagtatat acaaagtata acgatatggt
     2401 ggctactctc gaggccaggt gaaatatgag acttacg
//
