LOCUS       sequence-93259-e         713 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Donor vector for genomic targeting of a Tetracycline-inducible Cas9 
            cassette to the human AAVS1/PPP1R12C locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 713)
  AUTHORS   Gonzalez F, Zhu Z, Shi ZD, Lelli K, Verma N, Li QV, Huangfu D
  TITLE     An iCRISPR Platform for Rapid, Multiplexable, and Inducible Genome 
            Editing in Human Pluripotent Stem Cells.
  JOURNAL   Cell Stem Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 
            10.1016/j.stem.2014.05.018.
  PUBMED    24931489
REFERENCE   2  (bases 1 to 713)
  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 Stem 
            Cell. 2014 Jun 11. pii: S1934-5909(14)00205-7. doi: 
            10.1016/j.stem.2014.05.018."
COMMENT     Sequence Label: Puro-Cas9 donor
FEATURES             Location/Qualifiers
     source          1..713
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(259..279)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             complement(286..351)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     regulatory      351..360
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(385..409)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(429..453)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(450..488)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    500..518
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    536..554
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    571..589
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    643..661
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    678..696
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 gatccggttc ttccgtctgg tgtatcttct tctggcggtt ctcttcagcc gggtggcctc
       61 ggctgtttcg ccgctgtcga acagcagggc tccgatcagg ttcttcttga tgctgtgccg
      121 gtcggtgttg cccagcacct tgaatttctt gctgggcacc ttgtactcgt cggtgatcac
      181 ggcccagccc acagagttgg tgccgatgtc caggccgatg ctgtacttct tgtcggctgc
      241 tgggactccg tggataccga ccttccgctt cttctttggg gccatcttat cgtcatcgtc
      301 tttgtaatca atatcatgat ccttgtagtc tccgtcgtgg tccttatagt ccatggtggc
      361 accggtccaa cctgaaaacc gcggagcctg cttttttgta caaacttgtt gatcggaatt
      421 ctccaggcga tctgacggtt cactaaacga gctctgctta tataggcctc ccaccgtaca
      481 cgcctacctc gacatacgtt ctctatcact gatagggagt aaactcgaca tacgttctct
      541 atcactgata gggataaact cgacatacgt tctctatcac tgatagggag taaactcgac
      601 atacgttctc tatcactgat agggagtaaa ctcgacatac gttctctatc actgataggg
      661 agtaaactcg acatcgttct ctatcactga tagggagtaa actcgacata cgt
//
