LOCUS       sequence-217959-        6658 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses N-terminus to aa571 of PhiC31 integrase fused to FRB CID.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6658)
  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 6658)
  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: pBW1349_pCAG-PV1-PhiC31-N571-L1-FRB-NLS-BGHpA
FEATURES             Location/Qualifiers
     source          1..6658
                     /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"
     regulatory      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2432..2707
                     /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             2708..2728
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     polyA_signal    2738..2962
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3202..3221)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3267..3322
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3321..3340)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3683..3699)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3683..3699)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3696..3718)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3707..3723
                     /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(3731..3737,3738..3755,3756..3761))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3776..3797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      3882..4016
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4036..4170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4073..4092)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4127..4146
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4238..4255)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4409..4997)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4489..4508)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5168..5959,5960..6028))
                     /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     5791..5810
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6029..6133)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6164..6543
                     /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 gccaccatgg atacctacgc cggagcctac gacagacaga gccgggagag
      721 agagaacagc agcgccgcca gccccgccac ccagagaagc gccaacgagg ataaggccgc
      781 cgatctgcag agagaggtgg agagggacgg cggcagattc agatttgtgg gccacttcag
      841 cgaggcccct ggcaccagcg ccttcggcac cgccgagaga cccgagttcg agagaatcct
      901 gaacgagtgt agggccggca ggctgaacat gatcatcgtg tacgacgtgt cccggttcag
      961 caggctgaag gtgatggacg ccatccctat cgtgtccgag ctgctggccc tgggcgtgac
     1021 catcgtgtcc acccaggaag gcgtctttag acagggcaac gtgatggacc tgatccacct
     1081 gatcatgagg ctggacgcca gccacaagga gagcagcctg aagagcgcca agatcctgga
     1141 caccaagaac ctgcagaggg agctgggcgg ctatgtgggc ggcaaggccc cctacggctt
     1201 cgagctggtg tccgagacca aggagatcac ccggaacggc aggatggtga acgtggtgat
     1261 caacaagctg gcccacagca ccacccccct gaccggcccc ttcgagtttg agcccgacgt
     1321 gatcaggtgg tggtggcggg agatcaagac ccacaagcac ctgcctttca agcccggcag
     1381 ccaggccgcc atccaccccg gcagcatcac cggcctgtgt aagagaatgg acgccgacgc
     1441 cgtgcccacc agaggcgaga ccatcggcaa gaaaaccgcc agcagcgcct gggaccccgc
     1501 caccgtgatg agaatcctga gggaccctag gatcgccggc ttcgccgccg aggtgatcta
     1561 caagaagaag cccgacggca cccccaccac caagatcgag ggctacagaa tccagagaga
     1621 ccccatcacc ctgagacctg tggagctgga ctgtggccct atcatcgagc ctgccgagtg
     1681 gtacgagctg caggcctggc tggacggcag aggcagaggc aagggcctga gcagaggcca
     1741 ggccatcctg agcgccatgg acaagctgta ctgtgagtgt ggcgccgtga tgaccagcaa
     1801 gagaggcgag gagagcatca aggacagcta ccggtgccgg agaagaaagg tggtggaccc
     1861 cagcgcccct ggccagcacg agggcacctg taatgtgagc atggccgccc tggacaagtt
     1921 cgtggccgag cggatcttca acaagatccg gcacgccgag ggcgacgagg agaccctggc
     1981 cctgctgtgg gaggccgcca gaagattcgg caagctgacc gaggcccccg agaagagcgg
     2041 cgagagggcc aacctggtgg ccgagagagc cgacgccctg aacgccctgg aggagctgta
     2101 cgaggacaga gccgccggag cctatgacgg ccctgtgggc aggaagcact tcagaaagca
     2161 gcaggccgcc ctgaccctga gacagcaggg cgccgaggaa agactggccg agctggaggc
     2221 cgccgaggcc cctaagctgc ccctggatca gtggttcccc gaggatgccg acgccgaccc
     2281 caccggcccc aagtcctggt ggggcagagc cagcgtggac gacaagaggg tgttcgtggg
     2341 cctgttcgtg gataagatcg tggtgaccaa gagcaccacc ggcaggggct ccggagggtc
     2401 tggctccgga tcaagtggtg gcagcggtac catcctctgg catgagatgt ggcatgaagg
     2461 cctggaagag gcatctcgtt tgtactttgg ggaaaggaac gtgaaaggca tgtttgaggt
     2521 gctggagccc ttgcatgcta tgatggaacg gggcccccag actctgaagg aaacatcctt
     2581 taatcaggcc tatggtcgag atttaatgga ggcccaagag tggtgcagga agtacatgaa
     2641 atcagggaat gtcaaggacc tcctccaagc ctgggacctc tattatcatg tgttccgacg
     2701 aatctcaccc aagaaaaagc ggaaggtgtg agaattcctg tgccttctag ttgccagcca
     2761 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     2821 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     2881 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     2941 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3001 tatccccacg cgccctgtag cgtgtaccgc tgggagttcg tagacggaaa caaacgcaga
     3061 atccaagcgc ggccgcccag gatacataga ttaccacaac tccgagccct tccaccgcta
     3121 gcgcactcct caggtgcagg ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct
     3181 ggctcacaaa taccactgag atctttttcc ctctgccaaa aattatgggg acatcatgaa
     3241 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     3301 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     3361 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     3421 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     3481 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     3541 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     3601 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     3661 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3721 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3781 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3841 cgtgccagcg gatccgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     3901 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3961 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     4021 cttttgcaaa aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     4081 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4141 ctcatcaatg tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg
     4201 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4261 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4321 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4381 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4441 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4501 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4561 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     4621 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4681 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4741 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4801 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     4861 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4921 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4981 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5041 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5101 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5161 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5221 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5281 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5341 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5401 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5461 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5521 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     5581 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5641 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5701 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5761 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5821 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5881 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5941 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6001 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6061 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6121 taggggttcc gcgcacattt ccccgaaaag tgccacctgg gtcgacattg attattgact
     6181 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     6241 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     6301 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     6361 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     6421 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     6481 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     6541 atggtcgagg tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc
     6601 ccaattttgt atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggg
//
