LOCUS       sequence-217883-        4944 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Insert protein fragment into MluI/BspEI to fuse to FRB/NLS tags on 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4944)
  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 4944)
  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: pBW1308_pCAG-PV1-NTD-ENTRY-L1-FRB-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..4944
                     /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             718..993
                     /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             994..1014
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1024..1248
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1488..1507)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1553..1608
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1607..1626)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1969..1985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1969..1985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1982..2004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1993..2009
                     /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(2017..2023,2024..2041,2042..2047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2062..2083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2168..2302
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2322..2456
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2359..2378)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2413..2432
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2524..2541)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2695..3283)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2775..2794)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3454..4245,4246..4314))
                     /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     4077..4096
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4315..4419)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4450..4829
                     /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 cacgcgtaag tctactccgg agggtctggc tccggatcaa gtggtggcag cggtaccatc
      721 ctctggcatg agatgtggca tgaaggcctg gaagaggcat ctcgtttgta ctttggggaa
      781 aggaacgtga aaggcatgtt tgaggtgctg gagcccttgc atgctatgat ggaacggggc
      841 ccccagactc tgaaggaaac atcctttaat caggcctatg gtcgagattt aatggaggcc
      901 caagagtggt gcaggaagta catgaaatca gggaatgtca aggacctcct ccaagcctgg
      961 gacctctatt atcatgtgtt ccgacgaatc tcacccaaga aaaagcggaa ggtgtgagaa
     1021 ttcctgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1081 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1141 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1201 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     1261 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcgtg taccgctggg
     1321 agttcgtaga cggaaacaaa cgcagaatcc aagcgcggcc gcccaggata catagattac
     1381 cacaactccg agcccttcca ccgctagcgc actcctcagg tgcaggctgc ctatcagaag
     1441 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatct ttttccctct
     1501 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     1561 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     1621 tatgggaggg caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca
     1681 tatgcccata tgctggctgc catgaacaaa ggttggctat aaagaggtca tcagtatatg
     1741 aaacagcccc ctgctgtcca ttccttattc catagaaaag ccttgacttg aggttagatt
     1801 ttttttatat tttgttttgt gttatttttt tctttaacat ccctaaaatt ttccttacat
     1861 gttttactag ccagattttt cctcctctcc tgactactcc cagtcatagc tgtccctctt
     1921 ctcttatgga gatccctcga cctgcagccc aagcttggcg taatcatggt catagctgtt
     1981 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     2041 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     2101 gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc cgcatctcaa ttagtcagca
     2161 accatagtcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2221 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     2281 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt
     2341 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     2401 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct
     2461 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2521 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2581 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2641 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2701 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2761 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2821 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2881 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2941 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3001 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3061 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3121 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3181 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3241 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3301 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3361 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3421 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3481 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3541 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3601 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3661 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3721 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3781 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     3841 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     3901 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     3961 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4021 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4081 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4141 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4201 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4261 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4321 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4381 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4441 acctgggtcg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     4501 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     4561 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     4621 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     4681 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     4741 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     4801 tctacgtatt agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc
     4861 tccccatctc ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt
     4921 gtgcagcgat gggggcgggg gggg
//
