LOCUS       sequence-134126-         724 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A human codon-optimized SpCas9 fused to first 110 amino acids of 
            human GEMININ and chimeric guide RNA expression plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 724)
  AUTHORS   Gutschner T, Haemmerle M, Genovese G, Draetta GF, Chin L
  TITLE     Post-translational Regulation of Cas9 during G1 Enhances 
            Homology-Directed Repair.
  JOURNAL   Cell Rep. 2016 Feb 3. pii: S2211-1247(16)00040-1. doi: 
            10.1016/j.celrep.2016.01.019.
  PUBMED    26854237
REFERENCE   2  (bases 1 to 724)
  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 Rep. 
            2016 Feb 3. pii: S2211-1247(16)00040-1. doi: 
            10.1016/j.celrep.2016.01.019."
COMMENT     Sequence Label: pX330-U6-Chimeric_BB-CBh-hSpCas9-hGem(1/110)
FEATURES             Location/Qualifiers
     source          1..724
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             38..85
                     /codon_start=1
                     /product="bipartite nuclear localization signal from 
                     nucleoplasmin"
                     /label=nucleoplasmin NLS
                     /translation="KRPAATKKAGQAKKKK"
     CDS             98..427
                     /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"
     primer_bind     complement(461..478)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    467..674
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgagacacgg atcgacctgt ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa
       61 ggccggccag gcaaaaaaga aaaagggatc caagcttatg aatcccagta tgaagcagaa
      121 acaagaagaa atcaaagaga atataaagaa tagttctgtc ccaagaagaa ctctgaagat
      181 gattcagcct tctgcatctg gatctcttgt tggaagagaa aatgagctgt ccgcaggctt
      241 gtccaaaagg aaacatcgga atgaccactt aacatctaca acttccagcc ctggggttat
      301 tgtcccagaa tctagtgaaa ataaaaatct tggaggagtc acccaggagt catttgatct
      361 tatgattaaa gaaaatccat cctctcagta ttggaaggaa gtggcagaaa aacggagaaa
      421 ggcgctgtaa ctcgaggaat tcctagagct cgctgatcag cctcgactgt gccttctagt
      481 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
      541 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
      601 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agagaatagc
      661 aggcatgctg gggagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg
      721 cgcg
//
