LOCUS       sequence-167917-        3814 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR - FLIP targeting vector for mouse beta-catenin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3814)
  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 3814)
  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: pUC118-mCtnnb1-FLIP-Puro
FEATURES             Location/Qualifiers
     source          1..3814
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             37..72
                     /codon_start=1
                     /product="nanobody epitope tag corresponding to amino acids
                     16-27 of human beta-catenin (Traenkle et al., 2015)"
                     /label=BC2 tag
                     /translation="PDRKAAVSHWQQ"
     protein_bind    complement(889..922)
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(969..1002)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    complement(969..1002)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     primer_bind     complement(1028..1047)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1082..1101
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1371..1970)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1454..1474)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1951..1970
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    2104..2137
                     /label=lox5171
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTGTAC) (Shaw et al., 2021). lox5171 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        complement(2148..2647)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2196..2215)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2589..2612
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    2747..2780
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    2747..2780
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
ORIGIN
        1 acagctgacc tgatggagtt ggacatggcc atggagccgg acagaaaagc tgctgtcagc
       61 cactggcagc agcagtctta cttggattct ggaatccatt ctggtgccac caccacagct
      121 ccttccctga gtggcaaggg caaccctgag gaagaagatg ttgacacctc ccaagtcctt
      181 tatgaatggg agcaaggctt ttcccagtcc ttcacgcaag agcaagtagc tggtaaagca
      241 tttgtgtttg gaagctagca ttaagtttct tgacagggct gtgtgacagc tcagccacag
      301 cacaagtggg ttgaaggaag ggcggagggt agcggagtcc cacatgccta gtgagtgttg
      361 gatttacctt tttcagatat tgacgggcag tatgcaatga ctagggctca gagggtccga
      421 gctgccatgt tccctgagac gctagatgag ggcatgcaga tcccatccac gcagtttgac
      481 gctgctcatc ccactaatgt ccagcgcttg gctgaaccat cacagatgtt gaaacatgca
      541 gttgtcaatt tgattaacta tcaggatgac gcggaacttg ccacacgtgc aattcctgag
      601 ctgacaaaac tgctaaacga tgaggaccag gtaagcagtg atgtaacttg cttttcagtt
      661 tactttgagg gaagttctca aggggatgcc aatggctgcc tcacactaag ttttggaaac
      721 taatggtgcc tcttcatcct ttattctagg tggtagttaa taaagctgct gttatggtcc
      781 atcagctttc caaaaaggaa gcttccagac atgccatcat gcgctcccct cagatggtgt
      841 ctgccattgt acgcaccatg caggtaagta tcaaagatct ggcgcgccat aacttcgtat
      901 agtacacatt atacgaagtt atggctaagc tttgtgccat gccattgatg ctagcacagc
      961 ctgaatggat aacttcgtat aggatacctt atacgaagtt atgtcgacag acatgataag
     1021 atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg
     1081 tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaatt aacaagtgcc
     1141 atagagccca ccgcatcccc agcatgcctg ctattgtctt cccaatcctc ccccttgctg
     1201 tcctgcccca ccccaccccc cagaatagaa tgacacctac tcagacaatg cgatgcaatt
     1261 tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa ggaaggcacg
     1321 ggggaggggc aaacaacaga tggctggcaa ctagaaggca catcaggaat tcaggcaccg
     1381 ggcttgcggg tcatgcacca ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg
     1441 gtgaagccga gccgctcgta gaaggggagg ttgcggggcg cggaggtctc caggaaggcg
     1501 ggcaccccgg cgcgctcggc cgcctccact ccggggagca cgacggcgct gcccagaccc
     1561 ttgccctggt ggtcgggcga gacgccgacg gtggccagga accacgcggg ctccttgggc
     1621 cggtgcggcg ccaggaggcc ttccatctgt tgctgcgcgg ccagccggga accgctcaac
     1681 tcggccatgc gcgggccgat ctcggcgaac accgcccccg cttcgacgct ctccggcgtg
     1741 gtccagaccg ccaccgcggc gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg
     1801 acgcgcgtga ggaagagttc ttgcagctcg gtgacccgct cgatgtggcg gtccggatcg
     1861 acggtgtggc gcgtggcggg gtagtcggcg aacgcggcgg cgagggtgcg tacggccctg
     1921 gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt actcggtcat ggtacccacc
     1981 tgagggaggg aaaatagacc aataggcaga gagagtcagt gcctatcaga aacccaagag
     2041 tcttctctgt ctccacgtgc ccagtttcta ttggtctcct taaacctgtc ttgtaacctc
     2101 tagataactt cgtataatgt gtactatacg aagttatcga tggctgtcga aaggcccgga
     2161 gatgaggaag aggagaacag cgcggcagac gtgcgctttt gaagcgtgca gaatgccggg
     2221 cctccggagg accttcgggc gcccgccccg cccctgagcc cgcccctgag cccgcccccg
     2281 gacccacccc ttcccagcct ctgagcccag aaagcgaagg agcaaagctg ctattggccg
     2341 ctgccccaaa ggcctacccg cttccattgc tcagcggtgc tgtccatctg cacgagacta
     2401 gtgagacgtg ctacttccat ttgtcacgtc ctgcacgacg cgagctgcgg ggcggggggg
     2461 aacttcctga ctaggggagg agtagaaggt ggcgcgaagg ggccaccaaa gaacggagcc
     2521 ggttggcgcc taccggtgga tgtggaatgt gtgcgaggcc agaggccact tgtgtagcgc
     2581 caagtgccca gcggggctgc taaagcgcat gctccagact gccttgggaa aagcgcctcc
     2641 cctacccggg tgaggcggcc gcggttacaa gacaggttta aggagaccaa tagaaactgg
     2701 gcatgtggag acagagaaga ctcttgggtt tctgataggc actgacataa cttcgtataa
     2761 ggtatcctat acgaagttat tttccctccc ttagaataca aatgatgtag agacagctcg
     2821 ttgtactgct gggactctgc acaacctttc tcaccaccgc gagggcttgc tggccatctt
     2881 taagtctggt ggcatcccag cgctggtgaa aatgcttggg taagagaatg gccagggaac
     2941 ttgaagggta ggaagtagaa gggtgctggc atggctaaca aaggctctcc tcttcccagg
     3001 tcaccagtgg attctgtact gttctacgcc atcacgacac tgcataatct cctgctccat
     3061 caggaaggag ctaaaatggc agtgcgccta gctggtggac tgcagaaaat ggttgctttg
     3121 ctcaacaaaa caaacgtgaa attcttggct attacaacag actgccttca gatcttagct
     3181 tatggcaatc aagagagcaa ggtagagtga tgaaagttgt ttgtacagag tgtggagtta
     3241 ctaagttctt agtatgcatg tgcgcgcgcg cgcgcacaca cacacaggct caaagatttg
     3301 tgtctttaga cactgaatct gtgtgtacag tacggtgcta agcattgaag ggatggggag
     3361 caggcctcat aagtttacac ttgggcagag aggaggggct ggatagacag aggacatggt
     3421 gttttgaggt cttgtggcaa tctttgagaa gaagtgaggt gttacctagg aggaggcagt
     3481 gagaggatgc cgtccacagt cccgaggagg aagagggatg gtgtgctgga gacacagtgt
     3541 tagagttggg ctaaggctgg gccgggtgga tcctgtgagg ctttctttgt tggccatacc
     3601 aaagtccagg agataagatc ttaaacacag gcatggggtg agacctgcct atggaaatgt
     3661 tcaaggacct taagggtgag ggagtggata ttctgcagtg tctcctaagg caagtaacat
     3721 agaagctcag agtcagaagc taggggctct tagtgcccat tttcaactgg ctgtttcgac
     3781 agaactttga tcacttctac aagcaaccct gctt
//
