LOCUS       sequence-164430-        1018 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 1018)
  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 1018)
  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..1018
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    92..113
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(128..133,134..151,152..158)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    166..182
                     /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)."
     primer_bind     171..193
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     190..206
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     190..206
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        224..242
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     224..241
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     promoter        391..639
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     391..411
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     562..581
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        661..736
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     protein_bind    complement(748..772)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(847..866)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(848..866)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(873..890)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(873..889)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(882..904)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
       61 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
      121 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
      181 aatttcacac aggaaacagc tatgaccatg attacgccaa gctatttagg tgacactata
      241 gaatactcaa gctatgcatc aagcttggta ccgagctcgg atccactagt aacggccgcc
      301 agtgtgctgg aattcgccct ttgtacaaaa aagcaggctt taaaggaacc aattcagtcg
      361 actggatccg gtaccaaggt cgggcaggaa gagggcctat ttcccatgat tccttcatat
      421 ttgcatatac gatacaaggc tgttagagag ataattagaa ttaatttgac tgtaaacaca
      481 aagatattag tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt
      541 ttaaaattat gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat
      601 ttcttggctt tatatatctt gtggaaagga cgaaacaccg tacatcattt gtattctgca
      661 gttttagagc tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt
      721 ggcaccgagt cggtgctttt tttctagacc cagctttctt gtacaaagtt ggcattaaag
      781 ggcgaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
      841 aattcgccct atagtgagtc gtattacaat tcactggccg tcgttttaca acgtcgtgac
      901 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
      961 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctat
//
