LOCUS       sequence-217863-        4953 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Insert protein fragment into KpnI/EcoRI to fuse to FRB/NLS tag on 
            N-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4953)
  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 4953)
  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: pBW1050_pCAG-PV1-NLS-FRB-L1-CTD-ENTRY-BGHpA
FEATURES             Location/Qualifiers
     source          1..4953
                     /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             680..700
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             701..976
                     /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"
     polyA_signal    1033..1257
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1497..1516)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1562..1617
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1616..1635)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1978..1994)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1978..1994)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1991..2013)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2002..2018
                     /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(2026..2032,2033..2050,2051..2056))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2071..2092
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2177..2311
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2331..2465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2368..2387)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2422..2441
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2533..2550)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2704..3292)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2784..2803)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3463..4254,4255..4323))
                     /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     4086..4105
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4324..4428)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4459..4838
                     /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 gccaccatgc ccaagaaaaa gcggaaggtg atcctctggc atgagatgtg
      721 gcatgaaggc ctggaagagg catctcgttt gtactttggg gaaaggaacg tgaaaggcat
      781 gtttgaggtg ctggagccct tgcatgctat gatggaacgg ggcccccaga ctctgaagga
      841 aacatccttt aatcaggcct atggtcgaga tttaatggag gcccaagagt ggtgcaggaa
      901 gtacatgaaa tcagggaatg tcaaggacct cctccaagcc tgggacctct attatcatgt
      961 gttccgacga atctcatccg gagggtctgg ctccggatca agtggtggca gcggtaccaa
     1021 gtctacgaat tcctgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1081 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1141 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     1201 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     1261 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcgtgt
     1321 accgctggga gttcgtagac ggaaacaaac gcagaatcca agcgcggccg cccaggatac
     1381 atagattacc acaactccga gcccttccac cgctagcgca ctcctcaggt gcaggctgcc
     1441 tatcagaagg tggtggctgg tgtggccaat gccctggctc acaaatacca ctgagatctt
     1501 tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc
     1561 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     1621 ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt
     1681 ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat
     1741 cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga
     1801 ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt
     1861 tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct
     1921 gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc
     1981 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2041 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2101 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat
     2161 tagtcagcaa ccatagtccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     2221 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     2281 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct aacttgttta
     2341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     2401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     2461 ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2521 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     2581 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     2641 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     2701 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     2761 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     2821 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     2881 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     2941 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3001 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3061 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3121 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3181 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3241 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     3301 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     3361 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     3421 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     3481 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     3541 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     3601 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     3661 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     3721 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     3781 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     3841 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     3901 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     3961 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4021 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4081 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4141 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4201 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4261 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     4321 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     4381 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     4441 aaaagtgcca cctgggtcga cattgattat tgactagtta ttaatagtaa tcaattacgg
     4501 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     4561 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     4621 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     4681 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     4741 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     4801 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct
     4861 gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt
     4921 aattattttg tgcagcgatg ggggcggggg ggg
//
