LOCUS       sequence-95861-e         627 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9/gRNA Entry expression plasmid .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 627)
  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 627)
  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: piCRg Entry
FEATURES             Location/Qualifiers
     source          1..627
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     65..84
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        161..236
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(354..453)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(502..520)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(503..520)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(538..554)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(538..554)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(551..573)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    562..578
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(586..592,593..610,611..616))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
       61 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
      121 cttgtggaaa ggacgaaaca ccgggtcttc gagaagacct gttttagagc tagaaatagc
      181 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
      241 ttgttttaga gctagaaata gcaagttaaa ataaggctag tccgttttta gcgcgtgcgc
      301 caattctgca gacaaatggc tctagaggta cgaattcaag ggtgggcgcg ccgacccagc
      361 tttcttgtac aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca
      421 ggtcactatc agtcaaaata aaatcattat ttgccatcca gatccgagct cggtaccaag
      481 cttgatgcat agcttgagta ttctatagtg tcacctaaat agcttggcgt aatcatggtc
      541 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
      601 aagcataaag tgtaaagcct ggggtgc
//
