LOCUS       sequence-208326-        6709 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A S6 Kinase biosensor based on fluorescence resonance energy 
            transfer (FRET) and those based on bioluminescence energy transfer 
            (BRET).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6709)
  AUTHORS   Komatsu N, Terai K, Imanishi A, Kamioka Y, Sumiyama K, Jin T, Okada 
            Y, Nagai T, Matsuda M
  TITLE     A platform of BRET-FRET hybrid biosensors for optogenetics, chemical
            screening, and in vivo imaging.
  JOURNAL   Sci Rep. 2018 Jun 12;8(1):8984. doi: 10.1038/s41598-018-27174-x.
  PUBMED    29895862
REFERENCE   2  (bases 1 to 6709)
  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: "Sci Rep."; 
            date: "2018-06-12"; volume: "8(1):8984. doi"; pages: " 
            10.1038/s41598-018-27174-x"
COMMENT     Sequence Label: pCAGGS-6015nes
FEATURES             Location/Qualifiers
     source          1..6709
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1143..1490
                     /codon_start=1
                     /product="long flexible linker consisting of SAGG repeats 
                     (Komatsu et al., 2011)"
                     /label=EV linker
                     /translation="SAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGS
                     AGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGG
                     SAGGSAGGSAGG"
     CDS             3180..3206
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(3222..3241)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3287..3342
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3341..3360)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3703..3719)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3703..3719)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3716..3738)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3727..3743
                     /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(3751..3757,3758..3775,3776..3781))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3796..3817
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3876..4071
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3922..4057
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4077..4211
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4114..4133)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4168..4187
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4279..4296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4450..5038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4530..4549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5209..6000,6001..6069))
                     /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     5832..5851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6070..6174)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6209..6588
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
       61 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
      121 tgaccctgaa gcttctatgc accaccggca agctgcccgt gccctggccc accctcgtga
      181 ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
      241 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
      301 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
      361 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
      421 agtacaacta caacagccac aacgtctata tcaccgccga caagcagaag aacggcatca
      481 aggccaactt caagatccgc cacaacatcg aggacggcgg cgtgcagctc gccgaccact
      541 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
      601 gctaccagtc cgccctgttc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
      661 agttcctgac cgccgccggg atcactgagg gcatgaacga gctgtacctc gagaagtttt
      721 ctcaagaaca gatcggcgaa aacattgtgt gcagggtcat ttgtaccacg ggtcaaattc
      781 ccatccgaga tttgtcagct gatatttcac aagtgcttaa ggaaaaacga tccataaaga
      841 aagtttggac atttggtaga aacccagcct gtgactatca tttaggaaac atttcaagac
      901 tgtcaaataa gcatttccaa atactactag gagaagacgg taacctttta ttgaatgaca
      961 tttccactaa tgggacctgg ttaaatgggc aaaaagtcga gaagaacagc aatcagttac
     1021 tgtcccaagg tgatgaaata accgttggtg taggcgtgga atcagatatt ttatctctgg
     1081 tcattttcat aaacgacaaa tttaagcagt gcctggagca gaacaaagtt gatcgcggta
     1141 ccagtgctgg tggtagtgct ggtggtagtg ctggtggtag tgctggtggt agtgctggtg
     1201 gttccggcag tgctggtggt agtgctggtg gtagtaccag tgctggtggt agtgctggtg
     1261 gtagtgctgg tggtagtgct ggtggtagtg ctggtggttc cggcagtgct ggtggtagtg
     1321 ctggtggtag taccagtgct ggtggtagtg ctggtggtag tgctggtggt agtgctggtg
     1381 gtagtgctgg tggttccggc agtgctggtg gtagtgctgg tggtagtacc agtgctggtg
     1441 gtagtgctgg tggtagtgct ggtggtagtg ctggtggtag tgctggtggt tccggaaaca
     1501 ggcgaatcag aacacttacg gagcccgacg ttgattttaa tggcggccgc atggtgagca
     1561 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtca
     1621 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     1681 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1741 ccctgtcctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1801 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1861 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1921 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1981 acaactactt cagcgggaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     2041 ccaacttcaa gatccgccac aacatcgagg acggcggcgt gcagctcgcc gaccactacc
     2101 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2161 cccagtccgc cttaagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2221 tcttgaccgc cgccgggggt accaaggtgt acgaccccga gcagaggaag aggatgatca
     2281 ccggccccca gtggtgggcc aggtgcaagc agatgaacgt gctggacagc ttcatcaact
     2341 actacgacag cgagaagcac gccgagaacg ccgtgatctt cctgcacggc aacgccacta
     2401 gcagctacct gtggaggcac gtggtgcccc acatcgagcc cgtggccagg tgcatcatcc
     2461 ccgatctgat cggcatgggc aagagcggca agagcggcaa cggcagctac aggctgctgg
     2521 accactacaa gtacctgacc gcctggttcg agcttctgaa cctgcccaag aagatcatct
     2581 tcgtgggcca cgactggggc gccgccctgg ccttccacta cgcctacgag caccaggaca
     2641 ggatcaaggc catcgtgcac atggagagcg tggtggacgt gatcgagagc tgggacgagt
     2701 ggccagacat cgaggaggac atcgccctga tcaagagcga ggagggcgag aagatggtgc
     2761 tggagaacaa cttcttcgtg gagaccgtgc tgcccagcaa gatcatgaga aagctggagc
     2821 ccgaggagtt cgccgcctac ctggagccct tcaaggagaa gggcgaggtg agaagaccca
     2881 ccctgagctg gcccagagag atccccctgg tgaagggcgg caagcccgac gtggtgcaga
     2941 tcgtgagaaa ctacaacgcc tacctgagag ccagcgacga cctgcccaag ctgttcatcg
     3001 agggcgaccc cggcttcttc agcaacgcca tcgtggaggg cgccaagaag ttccccaaca
     3061 ccgagttcgt gaaggtgaag ggcctgcact tcctccagga ggacgccccc gacgagatgg
     3121 gcaagtacat caagagcttc gtggagagag tgctgaagaa cgagcagtct agactgcagc
     3181 tgcctcctct ggaacgcctg actctggatt aagtcgacgg atctttttcc ctctgccaaa
     3241 aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata aaggaaattt
     3301 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     3361 agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc aacatatgcc
     3421 catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta tatgaaacag
     3481 ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta gatttttttt
     3541 atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt acatgtttta
     3601 ctagccagat ttttcctcct ctcctgacta ctcccagtca tagctgtccc tcttctctta
     3661 tggagatccc tcgacctgca gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt
     3721 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     3781 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     3841 tttccagtcg ggaaacctgt cgtgccagcg gatccgcatc tcaattagtc agcaaccata
     3901 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3961 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     4021 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctaact
     4081 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4141 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4201 atgtctggat ccgctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     4261 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     4321 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     4381 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     4441 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     4501 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     4561 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     4621 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     4681 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4741 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4801 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4861 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4921 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4981 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     5041 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     5101 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     5161 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     5221 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     5281 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     5341 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     5401 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     5461 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     5521 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     5581 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     5641 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     5701 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     5761 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     5821 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     5881 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     5941 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6001 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6061 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     6121 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     6181 tccccgaaaa gtgccacctg ggtcgatcga cattgattat tgactagtta ttaatagtaa
     6241 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     6301 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     6361 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     6421 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     6481 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     6541 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc
     6601 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
     6661 tttatttttt aattattttg tgcagcgatg ggggcggggg ggggggggg
//
