LOCUS       Exported                7379 bp ds-DNA     circular SYN 23-AUG-2022
DEFINITION  Booster-PKA, a Red-Shifted Genetically Encoded FRET Biosensor for 
            monitoring PKA activity..
ACCESSION   .
VERSION     .
KEYWORDS    pCAGGS-4493NES
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7379)
  AUTHORS   Watabe T, Terai K, Sumiyama K, Matsuda M
  TITLE     Booster, a Red-Shifted Genetically Encoded Forster Resonance Energy 
            Transfer (FRET) Biosensor Compatible with Cyan Fluorescent 
            Protein/Yellow Fluorescent Protein-Based FRET Biosensors and Blue 
            Light-Responsive Optogenetic Tools.
  JOURNAL   ACS Sens. 2020 Feb 26. doi: 10.1021/acssensors.9b01941.
  PUBMED    32101394
REFERENCE   2  (bases 1 to 7379)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 23, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7379
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1606..1628
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1694..1713
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1743..1752
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1749..2444
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             3660..4313
                     /codon_start=1
                     /product="monomeric Kusabira-Orange 2 fluorescent protein"
                     /label=mKO2
                     /translation="MVSVIKPEMKMRYYMDGSVNGHEFTIEGEGTGRPYEGHQEMTLRV
                     TMAEGGPMPFAFDLVSHVFCYGHRVFTKYPEEIPDYFKQAFPEGLSWERSLEFEDGGSA
                     SVSAHISLRGNTFYHKSKFTGVNFPADGPIMQNQSVDWEPSTEKITASDGVLKGDVTMY
                     LKLEGGGNHKCQFKTTYKAAKEILEMPGDHYIGHRLVRKTEGNITEQVEDAVAHS"
     CDS             4359..4385
                     /codon_start=1
                     /product="nuclear export signal from the HIV Rev protein 
                     (Fischer et al., 1995)"
                     /label=NES
                     /translation="LPPLERLTL"
     primer_bind     complement(4401..4420)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4466..4521
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4520..4539)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4882..4898)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4882..4898)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4895..4917)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4906..4922
                     /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(4930..4960)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4975..4996
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5055..5250
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5101..5236
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5163..5182
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    5256..5390
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5293..5312)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5347..5366
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5458..5475)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5629..6217)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5709..5728)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6388..7248)
                     /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     7011..7030
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7249..7353)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggg gcgcgccrgg gggggggggg gggggggggg
      541 gggggggggg ggsgrggcgg rgaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
      721 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      781 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
      841 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
      901 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
      961 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
     1021 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
     1141 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
     1201 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     1261 gggcggggcc gcctcgggcc ggggrgggct cgggggaggg gcgcggcggc ccccggagcg
     1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1381 gggcgcaggg acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc
     1441 gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg
     1501 gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct
     1561 gtccgcgggg ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc
     1621 gtgtgaccgg cggctctagc tagagcctct gctaaccatg ttcatgcctt cttctttttc
     1681 ctacagctcc tgggcaacgt gctggttatt gtgctgtctc atcattttgg caagaattcg
     1741 ccgccaccat ggtgagcgag ctgattaagg agaacatgca catgaagctg tacatggagg
     1801 gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
     1861 gcacccagac catgagaatc aaggcggtcg agggcggccc tctccccttc gccttcgaca
     1921 tcctggctac cagcttcatg tacggcagca aaaccttcat caaccacacc cagggcatcc
     1981 ccgacttctt taagcagtcc ttccccgagg gcttcacatg ggagagagtc accacatacg
     2041 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
     2101 acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa
     2161 cactcggctg ggaggcctcc accgagaccc tgtaccccgc tgacggcggc ctggaaggca
     2221 gagccgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac ttgaagacca
     2281 catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tatgtggaca
     2341 gaagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg
     2401 ctgtggccag atactgcgac ctccctagca aactggggca cagactcgag aagttttctc
     2461 aagaacagat cggcgaaaac attgtgtgca gggtcatttg taccacgggt caaattccca
     2521 tccgagattt gtcagctgat atttcacaag tgcttaagga aaaacgatcc ataaagaaag
     2581 tttggacatt tggtagaaac ccagcctgtg actatcattt aggaaacatt tcaagactgt
     2641 caaataagca tttccaaata ctactaggag aagacggtaa ccttttattg aatgacattt
     2701 ccactaatgg gacctggtta aatgggcaaa aagtcgagaa gaacagcaat cagttactgt
     2761 cccaaggtga tgaaataacc gttggtgtag gcgtggaatc agatatttta tctctggtca
     2821 ttttcataaa cgacaaattt aagcagtgcc tggagcagaa caaagttgat cgcggtacca
     2881 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggtt
     2941 ccggcagtgc tggtggtagt gctggtggta gtaccagtgc tggtggtagt gctggtggta
     3001 gtgctggtgg tagtgctggt ggtagtgctg gtggttccgg cagtgctggt ggtagtgctg
     3061 gtggtagtac cagtgctggt ggtagtgctg gtggtagtgc tggtggtagt gctggtggta
     3121 gtgctggtgg ttccggcagt gctggtggta gtgctggtgg tagtaccagt gctggtggta
     3181 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggttcc ggcagtgctg
     3241 gtggtagtgc tggtggtagt accagtgctg gtggtagtgc tggtggtagt gctggtggta
     3301 gtgctggtgg tagtgctggt ggttccggca gtgctggtgg tagtgctggt ggtagtacca
     3361 gtgctggtgg tagtgctggt ggtagtgctg gtggtagtgc tggtggtagt accagtgctg
     3421 gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggttccggca
     3481 gtgctggtgg tagtgctggt ggtagtacca gtgctggtgg tagtgctggt ggtagtgctg
     3541 gtggtagtgc tggtggttcc ggcagtgctg gtggtagtgc tggtggtagt accagtgctg
     3601 gtggtagtgc tggtggtagt gctggtggta gtgctggtgg tagtgctggt ggttccggaa
     3661 tggtgagtgt gattaaacca gagatgaaga tgaggtacta catggacggc tccgtcaatg
     3721 ggcatgagtt cacaattgaa ggtgaaggca caggcagacc ttacgaggga catcaagaga
     3781 tgacactacg cgtcacaatg gccgagggcg ggccaatgcc tttcgcgttt gacttagtgt
     3841 cacacgtgtt ctgttacggc cacagagtat ttactaaata tccagaagag ataccagact
     3901 atttcaaaca agcatttcct gaaggcctgt catgggaaag gtcgttggag ttcgaagatg
     3961 gtgggtccgc ttcagtcagt gcgcatataa gccttagagg aaacaccttc taccacaaat
     4021 ccaaatttac tggggttaac tttcctgccg atggtcctat catgcaaaac caaagtgttg
     4081 attgggagcc atcaaccgag aaaattactg ccagcgacgg agttctgaag ggtgatgtta
     4141 cgatgtacct aaaacttgaa ggaggcggca atcacaaatg ccaattcaag actacttaca
     4201 aggcggcaaa agagattctt gaaatgccag gagaccatta catcggccat cgcctcgtca
     4261 ggaaaaccga aggcaacatt actgagcagg tagaagatgc agtagctcat tccgcggccg
     4321 cattgaggcg cgcgacgctg gttgactcta gactgcagct gcctcctctg gaacgcctga
     4381 ctctggatta agtcgacgga tctttttccc tctgccaaaa attatgggga catcatgaag
     4441 ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     4501 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttaaaacat
     4561 cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc tgccatgaac
     4621 aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt ccattcctta
     4681 ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt tgtgttattt
     4741 ttttctttaa catccctaaa attttcctta catgttttac tagccagatt tttcctcctc
     4801 tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct cgacctgcag
     4861 cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     4921 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     4981 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     5041 gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     5101 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     5161 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     5221 ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca gcttataatg
     5281 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5341 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc cgctgcatta
     5401 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5461 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5521 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5581 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5641 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5701 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5761 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5821 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5881 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5941 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6001 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6061 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6121 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6181 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6241 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6301 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6361 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6421 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6481 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6541 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6601 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6661 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6721 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6781 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6841 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6901 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6961 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7021 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7081 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7141 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7201 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7261 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7321 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctg
//
