LOCUS       sequence-216970-        6070 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses N-terminus to aa374 of Flp recombinase fused to FRB CID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6070)
  AUTHORS   Weinberg BH, Cho JH, Agarwal Y, Pham NTH, Caraballo LD, Walkosz M, 
            Ortega C, Trexler M, Tague N, Law B, Benman WKJ, Letendre J, Beal J,
            Wong WW
  TITLE     High-performance chemical- and light-inducible recombinases in 
            mammalian cells and mice.
  JOURNAL   Nat Commun. 2019 Oct 24;10(1):4845. doi: 10.1038/s41467-019-12800-7.
  PUBMED    31649244
REFERENCE   2  (bases 1 to 6070)
  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 
            Commun."; date: "2019-10-24"; volume: "10(1):4845. doi"; pages: " 
            10.1038/s41467-019-12800-7"
COMMENT     Sequence Label: pBW2280_pCAG-PV1-FlpO-N374-L1-FRB-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..6070
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     466..488
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     550..569
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1841..2116
                     /codon_start=1
                     /product="T2098L mutant of FKBP-rapamycin binding domain of
                     human FRAP"
                     /label=FRB*
                     /note="binds synthetic ligands such as AP21967"
                     /translation="ILWHEMWHEGLEEASRLYFGERNVKGMFEVLEPLHAMMERGPQTL
                     KETSFNQAYGRDLMEAQEWCRKYMKSGNVKDLLQAWDLYYHVFRRIS"
     CDS             2117..2137
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2147..2371
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2611..2630)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2676..2731
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2730..2749)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3092..3108)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3092..3108)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3105..3127)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3116..3132
                     /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)."
     promoter        complement(join(3140..3146,3147..3164,3165..3170))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3185..3206
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      3291..3425
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3445..3579
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3482..3501)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3536..3555
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3647..3664)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3818..4406)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3898..3917)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4577..5368,5369..5437))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5200..5219
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5438..5542)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5573..5952
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
       61 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
      121 gggcggggcc gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg
      181 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
      241 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
      301 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
      361 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
      421 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
      481 gtgtgaccgg cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac
      541 agctcctggg caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattgatta
      601 attcgagcga aggcgcgccc attactcgca tccattctca ggctgtctcg tctcgtctcg
      661 cacgcgtgcc gccaccatga gccagttcga catcctgtgc aagacccccc ccaaggtgct
      721 ggtgcggcag ttcgtggaga gattcgagag gcccagcggc gagaagatcg ccagctgtgc
      781 cgccgagctg acctacctgt gctggatgat cacccacaac ggcaccgcca tcaagagggc
      841 caccttcatg agctacaaca ccatcatcag caacagcctg agcttcgaca tcgtgaacaa
      901 gagcctgcag ttcaagtaca agacccagaa ggccaccatc ctggaggcca gcctgaagaa
      961 gctgatcccc gcctgggagt tcaccatcat cccttacaac ggccagaagc accagagcga
     1021 catcaccgac atcgtgtcca gcctgcagct gcagttcgag agcagcgagg aggccgacaa
     1081 gggcaacagc cacagcaaga agatgctgaa ggccctgctg tccgagggcg agagcatctg
     1141 ggagatcacc gagaagatcc tgaacagctt cgagtacacc agcaggttca ccaagaccaa
     1201 gaccctgtac cagttcctgt tcctggccac attcatcaac tgcggcaggt tcagcgacat
     1261 caagaacgtg gaccccaaga gcttcaagct ggtgcagaac aagtacctgg gcgtgatcat
     1321 tcagtgcctg gtgaccgaga ccaagacaag cgtgtccagg cacatctact ttttcagcgc
     1381 cagaggcagg atcgaccccc tggtgtacct ggacgagttc ctgaggaaca gcgagcccgt
     1441 gctgaagaga gtgaacagga ccggcaacag cagcagcaac aagcaggagt accagctgct
     1501 gaaggacaac ctggtgcgca gctacaacaa ggccctgaag aagaacgccc cctaccccat
     1561 cttcgctatc aagaacggcc ctaagagcca catcggcagg cacctgatga ccagctttct
     1621 gagcatgaag ggcctgaccg agctgacaaa cgtggtgggc aactggagcg acaagagggc
     1681 ctccgccgtg gccaggacca cctacaccca ccagatcacc gccatccccg accactactt
     1741 cgccctggtg tccaggtact acgcctacga ccccatcagc aaggagatga tcgccctgtc
     1801 cggagggtct ggctccggat caagtggtgg cagcggtacc atcctctggc atgagatgtg
     1861 gcatgaaggc ctggaagagg catctcgttt gtactttggg gaaaggaacg tgaaaggcat
     1921 gtttgaggtg ctggagccct tgcatgctat gatggaacgg ggcccccaga ctctgaagga
     1981 aacatccttt aatcaggcct atggtcgaga tttaatggag gcccaagagt ggtgcaggaa
     2041 gtacatgaaa tcagggaatg tcaaggacct cctccaagcc tgggacctct attatcatgt
     2101 gttccgacga atctcaccca agaaaaagcg gaaggtgtga gaattcctgt gccttctagt
     2161 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2221 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2281 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2341 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2401 tctagggggt atccccacgc gccctgtagc gtgtaccgct gggagttcgt agacggaaac
     2461 aaacgcagaa tccaagcgcg gccgcccagg atacatagat taccacaact ccgagccctt
     2521 ccaccgctag cgcactcctc aggtgcaggc tgcctatcag aaggtggtgg ctggtgtggc
     2581 caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa attatgggga
     2641 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     2701 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     2761 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     2821 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     2881 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     2941 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     3001 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     3061 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3121 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     3181 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     3241 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     3301 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     3361 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     3421 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa
     3481 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3541 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc
     3601 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3661 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3721 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3781 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3841 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3901 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3961 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4021 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4081 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4141 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4201 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4261 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4321 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4381 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4441 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4501 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4561 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4621 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4681 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4741 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4801 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4861 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4921 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4981 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5041 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5101 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5161 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5221 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5281 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5341 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5401 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5461 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5521 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg tcgacattga
     5581 ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg
     5641 gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc
     5701 cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat
     5761 tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat
     5821 catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat
     5881 gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt attagtcatc
     5941 gctattacca tggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc
     6001 tccccacccc caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg
     6061 gggggggggg
//
