LOCUS       sequence-164416-         884 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 884)
  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 884)
  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..884
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    88..109
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(124..129,130..147,148..154)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    162..178
                     /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     167..189
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     186..202
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     186..202
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(308..341)
                     /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(388..421)
                     /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(388..421)
                     /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(428..461)
                     /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(481..500)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     535..554
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
       61 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
      121 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
      181 tcacacagga aacagctatg accatgatta cgaattcgag ctcggtaccc ggggatcctc
      241 tagagtcgtg tgaagagcgc gatcgcgttt aaacgctctt cagtaagtat caaagatctg
      301 gcgcgccata acttcgtata gtacacatta tacgaagtta tggctaagct ttgtgccatg
      361 ccattgatgc tagcacagcc tgaatggata acttcgtata ggatacctta tacgaagtta
      421 tgtcgacgaa gttcctatac tatctagaga ataggaactt cagacatgat aagatacatt
      481 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
      541 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca attaacaagt gccatagagc
      601 ccaccgcatc cccagcatgc ctgctattgt cttcccaatc ctcccccttg ctgtcctgcc
      661 ccaccccacc ccccagaata gaatgacacc tactcagaca atgcgatgca atttcctcat
      721 tttattagga aaggacagtg ggagtggcac cttccagggt caaggaaggc acgggggagg
      781 ggcaaacaac agatggctgg caactagaag gcacatcagg aattcaggca ccgggcttgc
      841 gggtcatgca ccaggtgcgc ggtccttcgg gcacctcgac gtcg
//
