LOCUS       sequence-164434-         984 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  mCtnnb1 gRNA expressing vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 984)
  AUTHORS   Andersson-Rolf A, Mustata RC, Merenda A, Kim J, Perera S, Grego T, 
            Andrews K, Tremble K, Silva JC, Fink J, Skarnes WC, Koo BK
  TITLE     One-step generation of conditional and reversible gene knockouts.
  JOURNAL   Nat Methods. 2017 Jan 30. doi: 10.1038/nmeth.4156.
  PUBMED    28135257
REFERENCE   2  (bases 1 to 984)
  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: "Nat 
            Methods. 2017 Jan 30. doi: 10.1038/nmeth.4156."
COMMENT     Sequence Label: gRNA-mCtnnb1
FEATURES             Location/Qualifiers
     source          1..984
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    52..76
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_RNA        complement(88..163)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(185..433)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(243..262)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(413..433)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     promoter        complement(582..600)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(583..600)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(618..634)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(618..634)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(631..653)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    642..658
                     /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(666..672,673..690,691..696))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    711..732
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(849..866)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gctcgagcgg ccgccagtgt gatggatatc tgcagaattc gccctttaat gccaactttg
       61 tacaagaaag ctgggtctag aaaaaaagca ccgactcggt gccacttttt caagttgata
      121 acggactagc cttattttaa cttgctattt ctagctctaa aactgcagaa tacaaatgat
      181 gtacggtgtt tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa tactttcaag
      241 ttacggtaag catatgatag tccattttaa aacataattt taaaactgca aactacccaa
      301 gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat
      361 taattctaat tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg
      421 gaaataggcc ctcttcctgc ccgaccttgg taccggatcc agtcgactga attggttcct
      481 ttaaagcctg cttttttgta caaagggcga attccagcac actggcggcc gttactagtg
      541 gatccgagct cggtaccaag cttgatgcat agcttgagta ttctatagtg tcacctaaat
      601 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
      661 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
      721 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
      781 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
      841 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
      901 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
      961 atgtgagcaa aaggccagca aaag
//
