LOCUS       sequence-178929-        3006 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  HMEJ donor for fusing a p2A-mCherry-WPRE reporter to mouse Dbh. AAV 
            backbone..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3006)
  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 3006)
  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_Dbh HMEJ donor
FEATURES             Location/Qualifiers
     source          1..3006
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             824..880
                     /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             881..1591
                     /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(1026..1044)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1294..1313
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1592..2180
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1645..1665)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2063..2074)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 agaatagctt ctcacaaggt ggggaataaa aatagagcta tgggcagggg aggcaagaaa
       61 ccagcagtct atgacaagag ccccacagcg cagcttctcc ctccttatct gaggcctgct
      121 gggattcaaa ggcacctaat ttatttccca gtagagctag ccagcagcag cagtgccagc
      181 ccagagtgct ggtcagcaga ccccgcctgc tgtgttaggc aggggaggca ggcactgcag
      241 cagaaggtga acagaccttt cctggcagcg tttgggtacc ggattaattg ctaacccaaa
      301 cacccatgcc atggggagac ttgttcatcc cagagcagtt tgaggcaccg ccggatatgg
      361 ttagatcttg gcttttgcag acgccttgtt ggacagtggc taaacccacg gcctgcctcc
      421 aaagactctg atcctttcct cggagtatac agttgagtgc aggcatggag gtggagacgt
      481 ctggagggct tcaggttgct gacagagaac acaacagcct agggaaggga cgtggatcct
      541 ccggtgctaa ctctcaaagg ctgacttgga agaggtgtgg agtgttcagc atgggagaaa
      601 tgattttaag gacaagaatc caaccagtta gggagatagc agtttgcttg gcccctccag
      661 cctcggctta ccttctgctc cttccttaca gggtgagtgg aatctgcagc ctttgcctaa
      721 gatcacctcc acgctggaag aacccacccc acgctgcccc atccgacaga ctcagagccc
      781 cgctaacccc actgtgccca tcacaacaga ggcagatgct gaagccacta acttctccct
      841 gttgaaacaa gcaggggatg tcgaagagaa tcccgggcca atggtgagca agggcgagga
      901 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
      961 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     1021 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     1081 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     1141 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     1201 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     1261 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     1321 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     1381 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     1441 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     1501 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     1561 caccggcggc atggacgagc tgtacaagta aaatcaacct ctggattaca aaatttgtga
     1621 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     1681 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     1741 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     1801 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     1861 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     1921 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     1981 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     2041 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     2101 gctgccggct ctgcggcctc ttccgcgtct ccgccttcgc cctcagacga gtcggatctc
     2161 cctttgggcc gcctccccgc cacttgtgag aagctattct tcagaccctc ctcctcctgt
     2221 ccacctgggc tcacaccagc tctgtgtagc cctcccttcc tgtgtgaaga ctcacttcca
     2281 tagactagtg ctggccatcc agggagaagg agcatgtcac cttggagaca cttgcatgac
     2341 tgcaccccat gaggtccaac tgaacggaga agccctggac attatctaca cactgctgac
     2401 tccacatagg gacagctcgc tgtggtagtc cagattcaac accaccttgc ctccagaaca
     2461 gccttgccag gctgggtgca gactccccaa gccccacagt cccgaccctg ttggtcctct
     2521 ggtggtgact tgtgttagtg tatgccgtga caacactgct taaatattcc tttgcagagt
     2581 ggctcatgtt tcccagtggg cggcttccct gcaacaagac aggacaagtc gtttagcagt
     2641 tagagactca ccaggggaac tcgcttcgtg gcagagtaaa tagatatttt cgcccaccta
     2701 gaggaaaccc ccaacaagta gttccatgac ggagagccaa gatggctaga gccaggctct
     2761 gtagaggagt ggaacggcca gagctggttg cttctcctga tggtgacagg ccaggctctg
     2821 aaatagatag ttaggtgaag agctggcttg ggcccttaat gaaggagtga ctagagcaag
     2881 ccactcagct gcctgatctt cagtctcctt gtctgttaaa tgagacttac ccttacctag
     2941 cactaactgt gtacacatat gcatgtccag gattcctgcc ctgcccacct tgtgagaagc
     3001 tattct
//
