LOCUS       sequence-82217-e         992 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  An empty gRNA expression vector, used to create a gRNA to a specific
            sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 992)
  AUTHORS   Mali P, Yang L, Esvelt KM, Aach J, Guell M, Dicarlo JE, Norville JE,
            Church GM
  TITLE     RNA-Guided Human Genome Engineering via Cas9.
  JOURNAL   Science. 2013 Feb 15;339(6121):823-6. doi: 10.1126/science.1232033
  PUBMED    23287722
REFERENCE   2  (bases 1 to 992)
  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: "10.1126/science"; 
            journalName: "Science"; date: "2013-02-15- 15"; volume: "339"; 
            issue: "6121"; pages: "823-6"
COMMENT     Sequence Label: gRNA_Cloning Vector
FEATURES             Location/Qualifiers
     source          1..992
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     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(412..436)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        complement(521..539)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(522..539)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(557..573)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(557..573)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(570..592)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    581..597
                     /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(605..611,612..629,630..635))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    650..671
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(788..805)
                     /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 tcttaagtta aaataaggct
      361 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttcta gacccagctt
      421 tcttgtacaa agttggcatt aaagggcgaa ttccagcaca ctggcggccg ttactagtgg
      481 atccgagctc ggtaccaagc ttgatgcata gcttgagtat tctatagtgt cacctaaata
      541 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
      601 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
      661 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
      721 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
      781 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
      841 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
      901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
      961 ccataggctc cgcccccctg acgagcatca ca
//
