LOCUS       sequence-161617-        2020 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  a mock vector for generating FLIP-FlpE targeting vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2020)
  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 2020)
  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-FLIP-FlpE-Puro
FEATURES             Location/Qualifiers
     source          1..2020
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    complement(26..59)
                     /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(106..139)
                     /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(106..139)
                     /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(146..179)
                     /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(199..218)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     253..272
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(542..1141)
                     /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(625..645)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1122..1141
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    complement(1270..1303)
                     /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    1309..1342
                     /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(1353..1852)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1401..1420)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1794..1817
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    1860..1893
                     /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    1860..1893
                     /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 gtaagtatca aagatctggc gcgccataac ttcgtatagt acacattata cgaagttatg
       61 gctaagcttt gtgccatgcc attgatgcta gcacagcctg aatggataac ttcgtatagg
      121 ataccttata cgaagttatg tcgacgaagt tcctatacta tctagagaat aggaacttca
      181 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
      241 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
      301 taacaagtgc catagagccc accgcatccc cagcatgcct gctattgtct tcccaatcct
      361 cccccttgct gtcctgcccc accccacccc ccagaataga atgacaccta ctcagacaat
      421 gcgatgcaat ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca
      481 aggaaggcac gggggagggg caaacaacag atggctggca actagaaggc acatcaggaa
      541 ttcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt
      601 cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggaggtct
      661 ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc
      721 tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg
      781 gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg
      841 aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc
      901 tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac accttgccga
      961 tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc
     1021 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
     1081 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
     1141 tggtacccac ctgagggagg gaaaatagac caataggcag agagagtcag tgcctatcag
     1201 aaacccaaga gtcttctctg tctccacgtg cccagtttct attggtctcc ttaaacctgt
     1261 cttgtaaccg aagttcctat actatctaga gaataggaac ttctctagat aacttcgtat
     1321 aatgtgtact atacgaagtt atcgatggct gtcgaaaggc ccggagatga ggaagaggag
     1381 aacagcgcgg cagacgtgcg cttttgaagc gtgcagaatg ccgggcctcc ggaggacctt
     1441 cgggcgcccg ccccgcccct gagcccgccc ctgagcccgc ccccggaccc accccttccc
     1501 agcctctgag cccagaaagc gaaggagcaa agctgctatt ggccgctgcc ccaaaggcct
     1561 acccgcttcc attgctcagc ggtgctgtcc atctgcacga gactagtgag acgtgctact
     1621 tccatttgtc acgtcctgca cgacgcgagc tgcggggcgg gggggaactt cctgactagg
     1681 ggaggagtag aaggtggcgc gaaggggcca ccaaagaacg gagccggttg gcgcctaccg
     1741 gtggatgtgg aatgtgtgcg aggccagagg ccacttgtgt agcgccaagt gcccagcggg
     1801 gctgctaaag cgcatgctcc agactgcctt gggaaaagcg cctcccctac ccgggtgaga
     1861 taacttcgta taaggtatcc tatacgaagt tatgcggccg cggttacaag acaggtttaa
     1921 ggagaccaat agaaactggg catgtggaga cagagaagac tcttgggttt ctgataggca
     1981 ctgactctct ctgcctattg gtctattttc ccacccttag
//
