LOCUS       sequence-161615-        1931 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  a mock vector for generating FLIP targeting vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1931)
  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 1931)
  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-Puro
FEATURES             Location/Qualifiers
     source          1..1931
                     /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)."
     primer_bind     complement(165..184)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     219..238
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(508..1107)
                     /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(591..611)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     1088..1107
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     protein_bind    1241..1274
                     /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(1285..1784)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(1333..1352)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     1726..1749
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     protein_bind    1884..1917
                     /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    1884..1917
                     /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 tcgacagaca tgataagata cattgatgag tttggacaaa
      181 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
      241 tatttgtaac cattataagc tgcaattaac aagtgccata gagcccaccg catccccagc
      301 atgcctgcta ttgtcttccc aatcctcccc cttgctgtcc tgccccaccc caccccccag
      361 aatagaatga cacctactca gacaatgcga tgcaatttcc tcattttatt aggaaaggac
      421 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
      481 ctggcaacta gaaggcacat caggaattca ggcaccgggc ttgcgggtca tgcaccaggt
      541 gcgcggtcct tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa
      601 ggggaggttg cggggcgcgg aggtctccag gaaggcgggc accccggcgc gctcggccgc
      661 ctccactccg gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgagac
      721 gccgacggtg gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc
      781 catctgttgc tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc
      841 ggcgaacacc gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc
      901 gtcgtccgcg acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg
      961 cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta
     1021 gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag
     1081 gcgcaccgtg ggcttgtact cggtcatggt acccacctga gggagggaaa atagaccaat
     1141 aggcagagag agtcagtgcc tatcagaaac ccaagagtct tctctgtctc cacgtgccca
     1201 gtttctattg gtctccttaa acctgtcttg taacctctag ataacttcgt ataatgtgta
     1261 ctatacgaag ttatcgatgg ctgtcgaaag gcccggagat gaggaagagg agaacagcgc
     1321 ggcagacgtg cgcttttgaa gcgtgcagaa tgccgggcct ccggaggacc ttcgggcgcc
     1381 cgccccgccc ctgagcccgc ccctgagccc gcccccggac ccaccccttc ccagcctctg
     1441 agcccagaaa gcgaaggagc aaagctgcta ttggccgctg ccccaaaggc ctacccgctt
     1501 ccattgctca gcggtgctgt ccatctgcac gagactagtg agacgtgcta cttccatttg
     1561 tcacgtcctg cacgacgcga gctgcggggc gggggggaac ttcctgacta ggggaggagt
     1621 agaaggtggc gcgaaggggc caccaaagaa cggagccggt tggcgcctac cggtggatgt
     1681 ggaatgtgtg cgaggccaga ggccacttgt gtagcgccaa gtgcccagcg gggctgctaa
     1741 agcgcatgct ccagactgcc ttgggaaaag cgcctcccct acccgggtga ggcggccgcg
     1801 gttacaagac aggtttaagg agaccaatag aaactgggca tgtggagaca gagaagactc
     1861 ttgggtttct gataggcact gacataactt cgtataaggt atcctatacg aagttatttt
     1921 ccctccctta g
//
