LOCUS       sequence-246154-        6621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for Akt activity.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6621)
  AUTHORS   Maryu G, Matsuda M, Aoki K
  TITLE     Multiplexed Fluorescence Imaging of ERK and Akt Activities and 
            Cell-cycle Progression.
  JOURNAL   Cell Struct Funct. 2016 Jul 22;41(2):81-92. doi: 10.1247/csf.16007. 
            Epub 2016 May 28.
  PUBMED    27247077
REFERENCE   2  (bases 1 to 6621)
  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"; doi: "10.1247/csf.16007";
            journalName: "Cell Struct Funct"; date: "2016-07-22- 22"; volume: 
            "41"; issue: "2"; pages: "81-92"
COMMENT     Sequence Label: pCAGGS-Akt-FoxO3a-KTR-EGFP
FEATURES             Location/Qualifiers
     source          1..6621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          666..1686
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     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"
     CDS             join(2941..2943,2944..2946,2947..3459)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(2986..3007)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3247..3266)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3594..3615
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3643..3662)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3708..3763
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3762..3781)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4124..4140)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4124..4140)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4137..4159)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4148..4164
                     /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(4172..4178,4179..4196,4197..4202))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4217..4238
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4297..4492
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4343..4478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4498..4632
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4535..4554)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4589..4608
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4700..4717)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4871..5459)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4951..4970)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5630..6421,6422..6490))
                     /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     6253..6272
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6491..6595)
                     /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 gggggggggs gcgcgccmgg gggggggggg gggggggggg
      541 gggggggggg ggsgrggsgg rgrggkgcgg 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 cgggggrggg gcgcggcggc cccsggagcg
     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 caaagaattc
     1741 gccatggcag aggcaccggc ttccccggcc ccgctctctc cgctcgaagt ggagctggac
     1801 ccggagttcg agccccagag ccgtccgcga tcctgtacgt ggcccctgca aaggccggag
     1861 ctccaagcga gccctgccaa gccctcgggg gagacggccg ccgactccat gatccccgag
     1921 gaggaggacg atgaagacga cgaggacggc gggggacggg ccggctcggc catggcgatc
     1981 ggcggcggcg gcgggagcgg cacgctgggc tccgggctgc tccttgagga ctcggcccgg
     2041 gtgctggcac ccggagggca agaccccggg tctgggccag ccaccgcggc gggcgggctg
     2101 agcgggggta cacaggcgct gctgcagcct cagcaaccgc tgccaccgcc gcagccgggg
     2161 gtggctgggg gctccgggca gccgaggaaa tgttcgtcgc ggcggaacgc ctggggaaac
     2221 ctgtcctacg cggacctgat cacccgcgcc atcgagagct ccccggacaa acggctcact
     2281 ctgtcccaga tctacgagtg gatggtgcgt tgcgtgccct acttcaagga taagggcgac
     2341 agcaacagct ctgccggctg gaagaactcc atccggcgca acctgtcact gcatagtcga
     2401 ttcatgcggg tccagaatga gggaactggc aagagctctt ggtggatcat caaccctgat
     2461 ggggggaaga gcggaaaagc cccccggcgg cgggctgtct ccatggacaa tagcaacaag
     2521 tataccaaga gccgtggccg cgcagccaag aagaaggcag ccctgcagac agcccccgaa
     2581 tcagctgacg acagtccctc ccagctctcc aagtggcctg gcagccccac gtcacgcagc
     2641 agtgatgagc tggatgcgtg gacggacttc cgttcacgca ccaattctaa cgccagcaca
     2701 gtcagtggcc gcctgtcgcc catcatggca agcacagagt tggatgaagt ccaggacgat
     2761 gatgcgcctc tctcgcccat gctctacagc agctcagcca gcctgtcacc ttcagtaagc
     2821 aagccgtgca cggtggaact gccacggctg actgatatgg caggcaccat gaatctgaat
     2881 gatgggctga ctgaaaacct catggacgac ctgctggata acatcacgct cggcggccgc
     2941 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3001 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3061 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3121 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3181 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3241 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3301 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3361 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3421 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3481 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3541 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3601 ctgctggagt tcgtgaccgc cgcctctaga taagtcgacg gatctttttc cctctgccaa
     3661 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     3721 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     3781 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     3841 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     3901 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     3961 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     4021 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     4081 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     4141 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     4201 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     4261 ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat
     4321 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4381 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4441 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac
     4501 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     4561 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     4621 catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4681 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4741 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4801 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4861 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4921 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4981 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     5041 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     5101 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5161 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     5221 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5281 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5341 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5401 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5461 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5521 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5581 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5641 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5701 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5761 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5821 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5881 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5941 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6001 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6061 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6121 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6181 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6241 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6301 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6361 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6421 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6481 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6541 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6601 ttccccgaaa agtgccacct g
//
