LOCUS       sequence-167918-        3903 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  CRISPR - FLIP-FlpE targeting vector for mouse beta-catenin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 3903)
  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 3903)
  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-FlpE-Puro
FEATURES             Location/Qualifiers
     source          1..3903
                     /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)."
     protein_bind    complement(1009..1042)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(1062..1081)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1116..1135
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1405..2004)
                     /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(1488..1508)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1985..2004
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    complement(2133..2166)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    2172..2205
                     /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(2216..2715)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(2264..2283)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     2657..2680
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    2723..2756
                     /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    2723..2756
                     /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 atgtcgacga agttcctata
     1021 ctatctagag aataggaact tcagacatga taagatacat tgatgagttt ggacaaacca
     1081 caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct attgctttat
     1141 ttgtaaccat tataagctgc aattaacaag tgccatagag cccaccgcat ccccagcatg
     1201 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
     1261 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
     1321 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
     1381 gcaactagaa ggcacatcag gaattcaggc accgggcttg cgggtcatgc accaggtgcg
     1441 cggtccttcg ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg
     1501 gaggttgcgg ggcgcggagg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc
     1561 cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc
     1621 gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat
     1681 ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc
     1741 gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc
     1801 gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag
     1861 ctcggtgacc cgctcgatgt ggcggtccgg atcgacggtg tggcgcgtgg cggggtagtc
     1921 ggcgaacgcg gcggcgaggg tgcgtacggc cctggggacg tcgtcgcggg tggcgaggcg
     1981 caccgtgggc ttgtactcgg tcatggtacc cacctgaggg agggaaaata gaccaatagg
     2041 cagagagagt cagtgcctat cagaaaccca agagtcttct ctgtctccac gtgcccagtt
     2101 tctattggtc tccttaaacc tgtcttgtaa ccgaagttcc tatactatct agagaatagg
     2161 aacttctcta gataacttcg tataatgtgt actatacgaa gttatcgatg gctgtcgaaa
     2221 ggcccggaga tgaggaagag gagaacagcg cggcagacgt gcgcttttga agcgtgcaga
     2281 atgccgggcc tccggaggac cttcgggcgc ccgccccgcc cctgagcccg cccctgagcc
     2341 cgcccccgga cccacccctt cccagcctct gagcccagaa agcgaaggag caaagctgct
     2401 attggccgct gccccaaagg cctacccgct tccattgctc agcggtgctg tccatctgca
     2461 cgagactagt gagacgtgct acttccattt gtcacgtcct gcacgacgcg agctgcgggg
     2521 cgggggggaa cttcctgact aggggaggag tagaaggtgg cgcgaagggg ccaccaaaga
     2581 acggagccgg ttggcgccta ccggtggatg tggaatgtgt gcgaggccag aggccacttg
     2641 tgtagcgcca agtgcccagc ggggctgcta aagcgcatgc tccagactgc cttgggaaaa
     2701 gcgcctcccc tacccgggtg agataacttc gtataaggta tcctatacga agttatgcgg
     2761 ccgcggttac aagacaggtt taaggagacc aatagaaact gggcatgtgg agacagagaa
     2821 gactcttggg tttctgatag gcactgactc tctctgccta ttggtctatt ttcccaccct
     2881 tagaatacaa atgatgtaga gacagctcgt tgtactgctg ggactctgca caacctttct
     2941 caccaccgcg agggcttgct ggccatcttt aagtctggtg gcatcccagc gctggtgaaa
     3001 atgcttgggt aagagaatgg ccagggaact tgaagggtag gaagtagaag ggtgctggca
     3061 tggctaacaa aggctctcct cttcccaggt caccagtgga ttctgtactg ttctacgcca
     3121 tcacgacact acataatctc ctgctccatc aggaaggagc taaaatggca gtgcgcctag
     3181 ctggtggact gcagaaaatg gttgctttgc tcaacaaaac aaacgtgaaa ttcttggcta
     3241 ttacaacaga ctgccttcag atcttagctt atggcaatca agagagcaag gtagagtgat
     3301 gaaagttgtt tgtacagagt gtggagttac taagttctta gtatgcatgt gcgcgcgcgc
     3361 gcgcacacac acacaggctc aaagatttgt gtctttagac attgaatctg tgtgtacagt
     3421 acggtgctaa gcattgaagg gatggggagc aggcctcata agtttacact tgggcagaga
     3481 ggaggggctg gatagacaga ggacatggtg ttttgaggtc ttgtggcaat ctttgagaag
     3541 aagtgaggtg ttacctagga ggaggcagtg agaggatgcc gtccacagtc ccgaggagga
     3601 agagggatgg tgtgctggag acacagtgtt agagttgggc taaggctggg ccgggtggat
     3661 cctgtgaggc tttctttgtt ggccatacca aagtccagga gataagatct taaacacagg
     3721 catggggtga gacctgccta tggaaatgtt caaggacctt aagggtgagg gagtggatat
     3781 tctgcagtgt ctcctaaggc aagtaacata gaagctcaga gtcagaagct aggggctctt
     3841 agtgcccatt ttcaactggc tgtttcgaca gaactttgat cacttctaca agcaaccctg
     3901 ctt
//
