LOCUS       sequence-205981-        8250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent fusion protein for FoxO1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8250)
  AUTHORS   Gross SM, Dane MA, Bucher E, Heiser LM
  TITLE     Individual Cells Can Resolve Variations in Stimulus Intensity along 
            the IGF-PI3K-AKT Signaling Axis.
  JOURNAL   Cell Syst. 2019 Dec 18;9(6):580-588.e4. doi: 
            10.1016/j.cels.2019.11.005. Epub 2019 Dec 11.
  PUBMED    31838146
REFERENCE   2  (bases 1 to 8250)
  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: 
            "10.1016/j.cels.2019.11.005"; journalName: "Cell Syst"; date: 
            "2019-12-18- 18"; volume: "9"; issue: "6"; pages: "580-588.e4"
COMMENT     Sequence Label: pSBbi-FoxO1_1R_13A_3D
FEATURES             Location/Qualifiers
     source          1..8250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     525..545
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(1907..1928)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     2515..2536
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2590..2607)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2596..2820
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3252..3356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3357..3425,3426..4217)
                     /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     complement(3575..3594)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4388..4976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4877..4896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5130..5147
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5521..5540
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    5539..5594
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     5640..5659
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(5759..6358)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5842..5862)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6339..6358
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(6359..6415)
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(6727..6746)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     6996..7014
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(7154..7174)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        join(7645..8250,1..577)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(7876..8250,1..568)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
ORIGIN
        1 gggcggcgac ggggcccgtg cgtcccagcg cacatgttcg gcgaggcggg gcctgcgagc
       61 gcggccaccg agaatcggac gggggtagtc tcaagctggc cggcctgctc tggtgcctgg
      121 cctcgcgccg ccgtgtatcg ccccgccctg ggcggcaagg ctggcccggt cggcaccagt
      181 tgcgtgagcg gaaagatggc cgcttcccgg ccctgctgca gggagctcaa aatggaggac
      241 gcggcgctcg ggagagcggg cgggtgagtc acccacacaa aggaaaaggg cctttccgtc
      301 ctcagccgtc gcttcatgtg actccacgga gtaccgggcg ccgtccaggc acctcgatta
      361 gttctcgagc ttttggagta cgtcgtcttt aggttggggg gaggggtttt atgcgatgga
      421 gtttccccac actgagtggg tggagactga agttaggcca gcttggcact tgatgtaatt
      481 ctccttggaa tttgcccttt ttgagtttgg atcttggttc attctcaagc ctcagacagt
      541 ggttcaaagt ttttttcttc cattccaggt gtcgtgagga atttcgacat ttaaatttaa
      601 ttaaaccatg gcagaggctc cccaggtggt ggagaccgac ccggacttcg agccgctgcc
      661 ccggcagcgc tcctgtgctt ggccgctgcc caggccggag tttaaccagt ccaactcgac
      721 cacctccagt ccggcgccgt cgggcggagc ggccgccaac cccgacgccg cggcgagcct
      781 ggcctcggcg tccgctgtca gcaccgactt tatgagcaac ctgagcctgc tggaggagag
      841 tgaggacttc gcgcgggcgc caggctgcgt ggctgtggcg gcagcggctg cggccagcag
      901 aggcctgtgc ggggacttcc aaggtcctga ggcgggatgc gtacacccag cgcctccaca
      961 gcctccaccg accggaccgc tgtcgcagcc tccacccgtg cctccatccg ctgccgcagc
     1021 cgcggggcca ctcgcgggac agccgcgcaa gaccagctcg tcgcgccgca acgcgtgggg
     1081 caacctgtcg tacgccgacc tcatcaccaa ggccatcgag agttcagccg agaagaggct
     1141 caccctgtcg caaatctacg agtggatggt gaagagcgtg ccctacttca aggataaggg
     1201 cgacagcaac agctcggcgg gctggaagaa tgcaattcgc cgcaatctag cccttcacag
     1261 caagtttatt cgagtgcaga atgaaggaac tggaaagagt tcttggtgga tgctcaatcc
     1321 agagggaggc aagagcggaa aagctccccg gagaagagct gcggctatgg acaacaacag
     1381 taaatttgct aagagccgag ggcgggctgc taagaaaaaa gcatctctcc agtctgggca
     1441 agagggtcct ggagacgctc ctgggtctca gttttctaag tggcctgcgg ctcctggggc
     1501 tcacgctaac gatgactttg ataactggag tacatttcgt cctcgaacca gcgctaatgc
     1561 tagtactatc agtgggagag atgatgacat tatgacagag caggatgacc tgggagatgg
     1621 ggacgtgcat tccctggtgt atccaccctc tgctgccaag atggcatcta cgctgcccag
     1681 tctgtctgaa atcagcaatc cagaaaacat ggagaacctt ctggataatc tcaaccttct
     1741 ctcggctcca acatctttaa ctgtgtccac ccagtccgct cctggcagca tgatgcagca
     1801 ggcaccaagc ttcccacaca gtgtcaagac tacaacacac agctgggtgt caggaggatc
     1861 tatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1921 cggcgacgta aacggccaca agttcagcgt ccgcggcgag ggcgagggcg atgccaccaa
     1981 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2041 cctcgtgacc accttcggct acggcgtggc ctgcttcagc cgctaccccg accacatgaa
     2101 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctc
     2161 tttcaaggac gacggtacct acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2221 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2281 caagctggag tacaacttca acagccacta cgtctatatc acagccgaca agcagaagaa
     2341 cagcatcaag gctaacttca agatccgcca caacgttgag gacggcagcg tgcagctcgc
     2401 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2461 ctacctgagc catcagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     2521 cctgctggag ttcgtgaccg ctgccggaat cacactggga tccctcgagg tctagagctc
     2581 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2641 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2701 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     2761 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     2821 cttctgaggc ggaaagaacc agctgggcct actagatagc ggacccctta ccgaaacatc
     2881 gccgcattct gcagaggagt cgaacttagt gtatgtaaac ttctgaccca ctggaattgt
     2941 gatacagtga attataagtg aaataatctg tctgtaaaca attgttggaa aaatgacttg
     3001 tgtcatgcac aaagtagatg tcctaactga cttgccaaaa ctattgtttg ttaacaagaa
     3061 atttgtggag tagttgaaaa acgagtttta atgactccaa cttaagtgta tgtaaacttc
     3121 cgacttcaac tgtataggga tccaatttta acaaaatatt aacgcttaca atttcctgat
     3181 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatcaggt ggcacttttc
     3241 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3301 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3361 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     3421 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     3481 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     3541 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     3601 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     3661 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     3721 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     3781 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     3841 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     3901 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     3961 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4021 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4081 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4141 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4201 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4261 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4321 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4381 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4441 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4501 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4561 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4621 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4681 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4741 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4801 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4861 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4921 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4981 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5041 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5101 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggatccct
     5161 atacagttga agtcggaagt ttacatacac cttagccaaa tacatttaaa ctcacttttt
     5221 cacaattcct gacatttaat cctagtaaaa attccctgtc ttaggtcagt taggatcacc
     5281 actttatttt aagaatgtga aatatcagaa taatagtaga gagaatgatt catttcagct
     5341 tttatttctt tcatcacatt cccagtgggt cagaagttta catacactcg actcctctgc
     5401 agaatgcggc gatgtttcgg taaggggtcc cgagtagcta gttcatggca gccagcatat
     5461 ggcatatgtt gccaaactct aaaccaaata ctcattctga tgttttaaat gatttgccct
     5521 cccatatgtc cttccgagtg agagacacaa aaaattccaa cacactattg caatgaaaat
     5581 aaatttcctt tattagccag aagtcagatg ctcaaggggc ttcatgatgt ccccataatt
     5641 tttggcagag ggaaaaagat ctcagtggta tttgtgagcc agggcattgg ccacaccagc
     5701 caccaccttc tgataggcag cctgcacctg aggagtgaat tcacgacagg ccttcgaatc
     5761 aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc ttcgggcacc tcgacgtcgg
     5821 cggtgacggt gaagccgagc cgctcgtaga aggggaggtt gcggggcgcg gaggtctcca
     5881 ggaaggcggg caccccggcg cgctcggccg cctccactcc ggggagcacg acggcgctgc
     5941 ccagaccctt gccctggtgg tcgggcgaga cgccgacggt ggccaggaac cacgcgggct
     6001 ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg ctgcgcggcc agccgggaac
     6061 cgctcaactc ggccatgcgc gggccgatct cggcgaacac cgcccccgct tcgacgctct
     6121 ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc gacccacacc ttgccgatgt
     6181 cgagcccgac gcgcgtgagg aagagttctt gcagctcggt gacccgctcg atgtggcggt
     6241 ccggatcgac ggtgtggcgc gtggcggggt agtcggcgaa cgcggcggcg agggtgcgta
     6301 cggccctggg gacgtcgtcg cgggtggcga ggcgcaccgt gggcttgtac tcggtcatgg
     6361 ggccaggatt ctcctcgacg tcaccagcct gcttcagcag gctgaagtta gtagctccac
     6421 tgcccttgta cagatccagt ttcgcacgct tagctgcagg ctcgtccatg ccgccggtgg
     6481 agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg
     6541 tgatgtccaa cttgatgtcg acgttgtagg cgccgggcag ctgcacgggc ttcttggcct
     6601 tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct
     6661 tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc
     6721 ccatggtctt cttctgcatt acggggccgt cggaggggaa gttggtgccg cgcagcttca
     6781 ccttgtagat gaactcgccg tcctgcaggg aggagtcctg ggtcacggtc accacgccgc
     6841 cgtcctcgaa gttcatcacg cgctcccact tgaagccccc ggggaaggac agcttcaagt
     6901 agtcggggat gtcggcgggg tgcttcacgt aggccttgga gccgtacatg aactgagggg
     6961 acaggatgtc ccaggcgaag ggcagggggc cacccttggt caccttcagc ttggcggtct
     7021 gggtgccctc gtaggggcgg ccctcgccct cgccctcgat ctcgaactcg tggccgttca
     7081 cggagccctc catgtgcacc ttgaagcgca tgaactcctt gatgatggcc atgttatcct
     7141 cctcgccctt gctcaccttc cgctttttct taggactcat ggtggcacta gtgtctacgg
     7201 ctatatacct tgaaacaggg gcggcagaag tgacgcaacg cggttgcgcg gacggcttcc
     7261 tgtgttttca ttggtcgaag cgttctgcgc gtgcgcagag attccccagg gaaggcgggc
     7321 cctcttccga atgtccaatc cggacattct tccggttgga cctccgcggt cagcgctgtg
     7381 cccggcgaga gatcacgtgg ggcgcggagg cggtgctgct ggggcacggc cgtccagcct
     7441 cggcggccat atttttgagg ggctgttcat ctcgttcaca cgctctgtcc gccatgtttg
     7501 tgagtggaag cgccattacc ccttcaagcg actgaaggct gcagggcctc tggtggcccg
     7561 catggggaga ccagacccgc caggcccgcc tttccgcact cagtccgggc ttactttatt
     7621 ttgtgagaca gggtctcgcc tagaggctcc ggtgcccgtc agtgggcaga gcgcacatcg
     7681 cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg
     7741 tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt
     7801 gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg caacgggttt
     7861 gccgccagaa cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt
     7921 tatggccctt gcgtgccttg aattacttcc acgcccctgg ctgcagtacg tgattcttga
     7981 tcccgagctt cgggttggaa gtgggtggga gagttcgagg ccttgcgctt aaggagcccc
     8041 ttcgcctcgt gcttgagttg aggcctggct tgggcgctgg ggccgccgcg tgcgaatctg
     8101 gtggcacctt cgcgcctgtc tcgctgcttt cgataagtct ctagccattt aaaatttttg
     8161 atgacctgct gcgacgcttt ttttctggca agatagtctt gtaaatgcgg gccaagatct
     8221 gcacactggt atttcggttt ttggggccgc
//
