LOCUS       sequence-122673-         922 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9 sgRNA vector for cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 922)
  AUTHORS   Chen Y, Cao J, Xiong M, Petersen AJ, Dong Y, Tao Y, Huang CT, Du Z, 
            Zhang SC
  TITLE     Engineering Human Stem Cell Lines with Inducible Gene Knockout using
            CRISPR/Cas9.
  JOURNAL   Cell Stem Cell. 2015 Jul 1. pii: S1934-5909(15)00261-1. doi: 
            10.1016/j.stem.2015.06.001.
  PUBMED    26145478
REFERENCE   2  (bases 1 to 922)
  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. 2015 Jul 1. pii: S1934-5909(15)00261-1. doi: 
            10.1016/j.stem.2015.06.001."
COMMENT     Sequence Label: Cas9 sgRNA vector
FEATURES             Location/Qualifiers
     source          1..922
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        115..363
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     115..135
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     286..305
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    complement(449..473)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(558..576)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(559..576)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(594..610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(594..610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(607..629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    618..634
                     /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(642..648,649..666,667..672))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    687..708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(825..842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctcgagcggc cgccagtgtg atggatatct gcagaattcg ccctttgtac aaaaaagcag
       61 gctttaaagg aaccaattca gtcgactgga tccggtacca aggtcgggca ggaagagggc
      121 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
      181 agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
      241 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
      301 accgtaactt gaaagtattt cgatttcttg gctttatata tcttgtggaa aggacgggtc
      361 ttcgagaaga cgtgaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
      421 tggcaccgag tcggtgcttt ttttctagac ccagctttct tgtacaaagt tggcattaaa
      481 gggcgaattc cagcacactg gcggccgtta ctagtggatc cgagctcggt accaagcttg
      541 atgcatagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc atggtcatag
      601 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
      661 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
      721 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
      781 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
      841 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
      901 ttatccacag aatcagggga ta
//
