LOCUS       sequence-156759-         937 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Used for in vitro transcription of SpCas9-Gem mRNA. Cas9 is fused to
            Geminin peptide and is degraded during G0/G1 phases of the 
            cell-cycle to minimize indels caused by non-homologous end joining.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 937)
  AUTHORS   Howden SE, McColl B, Glaser A, Vadolas J, Petrou S, Little MH, 
            Elefanty AG, Stanley EG
  TITLE     A Cas9 Variant for Efficient Generation of Indel-Free Knockin or 
            Gene-Corrected Human Pluripotent Stem Cells.
  JOURNAL   Stem Cell Reports. 2016 Sep 13;7(3):508-17. doi: 
            10.1016/j.stemcr.2016.07.001. Epub 2016 Aug 4.
  PUBMED    27499201
REFERENCE   2  (bases 1 to 937)
  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"; doi: 
            "10.1016/j.stemcr.2016.07.001"; journalName: "Stem Cell Reports"; 
            date: "2016-09-13- 13"; volume: "7"; issue: "3"; pages: "508-17"
COMMENT     Sequence Label: pDNR-SpCas9-Gem
FEATURES             Location/Qualifiers
     source          1..937
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(30..359)
                     /codon_start=1
                     /product="degron consisting of residues 1-110 of human 
                     Geminin (Zielke and Edgar, 2015)"
                     /label=Gem1 (1-110)
                     /translation="MNPSMKQKQEEIKENIKNSSVPRRTLKMIQPSASGSLVGRENELS
                     AGLSKRKHRNDHLTSTTSSPGVIVPESSENKNLGGVTQESFDLMIKENPSSQYWKEVAE
                     KRRKAL"
ORIGIN
        1 gtttaaactc attactaacc ggtagattac agcgcctttc tccgtttttc tgccacttcc
       61 ttccaatact gagaggatgg attttcttta atcataagat caaatgactc ctgggtgact
      121 cctccaagat ttttattttc actagattct gggacaataa ccccagggct ggaagttgta
      181 gatgttaagt ggtcattccg atgtttcctt ttggacaagc ctgcggacag ctcattctct
      241 cttccaacaa gagatccaga tgcagaaggc tgaatcatct tcagagttct tcttgggaca
      301 gaactattct ttatattctc tttgatttct tcttgtttct gcttcatact gggattcatc
      361 tcgagcggcc gccagtgtga tggatatccc ttgtcagccc tgctgtctcc accgagctga
      421 gagaggtcga ttcttgtttc atagagcccc gtaattgact gatgaatcag tgtggcgtcc
      481 aggacctcct ttgtagaggt gtaccgcttt ctgtctatgg tggtgtcgaa gtacttgaag
      541 gctgcaggcg cgcccaagtt ggtcagagta aacaagtgga taatgttttc tgcctgctcc
      601 ctgatgggct tatccctgtg cttattgtaa gcagaaagca ccttatcgag gttagcgtcg
      661 gcgaggatca ctcttttgga gaattcgctt atttgctcga tgatctcatc aaggtagtgt
      721 ttgtgttgtt ccacgaacag ctgcttctgc tcattatctt cgggagaccc tttgagcttt
      781 tcatagtggc tggccagata caagaaatta acgtatttag agggcagtgc cagctcgtta
      841 cctttctgca gctcgcccgc actagcgagc attcgtttcc ggccgttttc aagctcaaag
      901 agagagtact tgggaagctt aatgatgagg tcttttt
//
