LOCUS       sequence-217862-        4993 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Insert protein fragment into MluI/BspEI to fuse to FKBP/NLS tags on 
            C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4993)
  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 4993)
  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: pBW1049_pCAG-PV1-NTD-ENTRY-L1-FKBP-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..4993
                     /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             724..1044
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1045..1065
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    1075..1299
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1539..1558)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1604..1659
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1658..1677)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2020..2036)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2020..2036)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2033..2055)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2044..2060
                     /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(2068..2074,2075..2092,2093..2098))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      2219..2353
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2373..2507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2410..2429)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2464..2483
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2575..2592)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2746..3334)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2826..2845)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3505..4296,4297..4365))
                     /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     4128..4147
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4366..4470)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4501..4880
                     /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 cggtacctct
      721 agaggagtgc aggtggaaac catctcccca ggagacgggc gcaccttccc caagcgcggc
      781 cagacctgcg tggtgcacta caccgggatg cttgaagatg gaaagaaatt tgattcctcc
      841 cgggacagaa acaagccctt taagtttatg ctaggcaagc aggaggtgat ccgaggctgg
      901 gaagaagggg ttgcccagat gagtgtgggt cagagagcca aactgactat atctccagat
      961 tatgcctatg gtgccactgg gcacccaggc atcatcccac cacatgccac tctcgtcttc
     1021 gatgtggagc ttctaaaact ggaacccaag aaaaagcgga aggtgtgaga attcctgtgc
     1081 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1141 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1201 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     1261 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     1321 gctggggctc tagggggtat ccccacgcgc cctgtagcgt gtaccgctgg gagttcgtag
     1381 acggaaacaa acgcagaatc caagcgcggc cgcccaggat acatagatta ccacaactcc
     1441 gagcccttcc accgctagcg cactcctcag gtgcaggctg cctatcagaa ggtggtggct
     1501 ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc tgccaaaaat
     1561 tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag gaaatttatt
     1621 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     1681 gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac atatgcccat
     1741 atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat gaaacagccc
     1801 cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat tttttttata
     1861 ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca tgttttacta
     1921 gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct tctcttatgg
     1981 agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     2041 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     2101 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     2161 ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc aaccatagtc
     2221 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     2281 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     2341 ttttttggag gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt
     2401 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     2461 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg
     2521 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     2581 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     2641 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2701 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2761 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2821 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2881 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2941 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3001 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3061 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3121 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3181 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3241 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3301 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3361 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3421 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3481 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     3541 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     3601 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     3661 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     3721 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     3781 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     3841 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     3901 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     3961 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4021 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4081 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4141 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     4201 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     4261 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     4321 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     4381 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4441 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc
     4501 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     4561 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     4621 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     4681 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     4741 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     4801 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
     4861 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
     4921 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
     4981 tgggggcggg ggg
//
