LOCUS       sequence-201199-        8253 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 8253)
  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 8253)
  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..8253
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        109..213
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(214..282,283..1074)
                     /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(432..451)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1245..1833
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1734..1753
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1987..2004
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2378..2397
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    2396..2451
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     2497..2516
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(2616..3215)
                     /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(2699..2719)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     3196..3215
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(3216..3272)
                     /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(3584..3603)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     3853..3871
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(4011..4031)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        4502..5684
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          4733..5675
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     5632..5652
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(7014..7035)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     7622..7643
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7700..7717)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7706..7930
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 aattttaaca aaatattaac gcttacaatt tcctgatgcg gtattttctc cttacgcatc
       61 tgtgcggtat ttcacaccgc atcaggtggc acttttcggg gaaatgtgcg cggaacccct
      121 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
      181 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
      241 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
      301 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
      361 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
      421 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
      481 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
      541 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
      601 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
      661 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
      721 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
      781 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
      841 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
      901 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
      961 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     1021 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     1081 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1141 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1201 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     1261 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     1321 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     1381 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1441 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1501 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1561 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1621 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1681 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1741 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     1801 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     1861 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     1921 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     1981 gagcgcagcg agtcagtgag cgaggaagcg gatccctata cagttgaagt cggaagttta
     2041 catacacctt agccaaatac atttaaactc actttttcac aattcctgac atttaatcct
     2101 agtaaaaatt ccctgtctta ggtcagttag gatcaccact ttattttaag aatgtgaaat
     2161 atcagaataa tagtagagag aatgattcat ttcagctttt atttctttca tcacattccc
     2221 agtgggtcag aagtttacat acactcgact cctctgcaga atgcggcgat gtttcggtaa
     2281 ggggtcccga gtagctagtt catggcagcc agcatatggc atatgttgcc aaactctaaa
     2341 ccaaatactc attctgatgt tttaaatgat ttgccctccc atatgtcctt ccgagtgaga
     2401 gacacaaaaa attccaacac actattgcaa tgaaaataaa tttcctttat tagccagaag
     2461 tcagatgctc aaggggcttc atgatgtccc cataattttt ggcagaggga aaaagatctc
     2521 agtggtattt gtgagccagg gcattggcca caccagccac caccttctga taggcagcct
     2581 gcacctgagg agtgaattca cgacaggcct tcgaatcagg caccgggctt gcgggtcatg
     2641 caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc
     2701 tcgtagaagg ggaggttgcg gggcgcggag gtctccagga aggcgggcac cccggcgcgc
     2761 tcggccgcct ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg
     2821 ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg cggcgccagg
     2881 aggccttcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc catgcgcggg
     2941 ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca gaccgccacc
     3001 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag
     3061 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg
     3121 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg
     3181 gtggcgaggc gcaccgtggg cttgtactcg gtcatggggc caggattctc ctcgacgtca
     3241 ccagcctgct tcagcaggct gaagttagta gctccactgc ccttgtacag atccagttta
     3301 gcacgcttag ctgcaggctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg
     3361 tactgttcca cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg
     3421 ttgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg
     3481 tcgtagtggc cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg
     3541 tcctcggggt acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg
     3601 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc
     3661 tgcagggagg agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc
     3721 tcccacttga agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc
     3781 ttcacgtagg ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
     3841 agggggccac ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc
     3901 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
     3961 aagcgcatga actccttgat gatggccatg ttatcctcct cgcccttgct caccttccgc
     4021 tttttcttag gactcatggt ggcactagtg tctacggcta tataccttga aacaggggcg
     4081 gcagaagtga cgcaacgcgg ttgcgcggac ggcttcctgt gttttcattg gtcgaagcgt
     4141 tctgcgcgtg cgcagagatt ccccagggaa ggcgggccct cttccgaatg tccaatccgg
     4201 acattcttcc ggttggacct ccgcggtcag cgctgtgccc ggcgagagat cacgtggggc
     4261 gcggaggcgg tgctgctggg gcacggccgt ccagcctcgg cggccatatt tttgaggggc
     4321 tgttcatctc gttcacacgc tctgtccgcc atgtttgtga gtggaagcgc cattacccct
     4381 tcaagcgact gaaggctgca gggcctctgg tggcccgcat ggggagacca gacccgccag
     4441 gcccgccttt ccgcactcag tccgggctta ctttattttg tgagacaggg tctcgcctag
     4501 aggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
     4561 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
     4621 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
     4681 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
     4741 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
     4801 tacttccacg cccctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg
     4861 ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg
     4921 cttggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg
     4981 ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt
     5041 tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg
     5101 gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc
     5161 tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg
     5221 tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg
     5281 caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat
     5341 ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct
     5401 ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc
     5461 tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg
     5521 cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga
     5581 tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc
     5641 agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaggaatt tcgacattta
     5701 aatttaatta aaccatggca gaggctcccc aggtggtgga gaccgacccg gacttcgagc
     5761 cgctgccccg gcagcgctcc tgtgcttggc cgctgcccag gccggagttt aaccagtcca
     5821 actcgaccac ctccagtccg gcgccgtcgg gcggagcggc cgccaacccc gacgccgcgg
     5881 cgagcctggc ctcggcgtcc gctgtcagca ccgactttat gagcaacctg agcctgctgg
     5941 aggagagtga ggacttcgcg cgggcgccag gctgcgtggc tgtggcggca gcggctgcgg
     6001 ccagcagagg cctgtgcggg gacttccaag gtcctgaggc gggatgcgta cacccagcgc
     6061 ctccacagcc tccaccgacc ggaccgctgt cgcagcctcc acccgtgcct ccatccgctg
     6121 ccgcagccgc ggggccactc gcgggacagc cgcgcaagac cagctcgtcg cgccgcaacg
     6181 cgtggggcaa cctgtcgtac gccgacctca tcaccaaggc catcgagagt tcagccgaga
     6241 agaggctcac cctgtcgcaa atctacgagt ggatggtgaa gagcgtgccc tacttcaagg
     6301 ataagggcga cagcaacagc tcggcgggct ggaagaatgc aattcgccgc aatctagccc
     6361 ttcacagcaa gtttattcga gtgcagaatg aaggaactgg aaagagttct tggtggatgc
     6421 tcaatccaga gggaggcaag agcggaaaag ctccccggag aagagctgcg gctatggaca
     6481 acaacagtaa atttgctaag agccgagggc gggctgctaa gaaaaaagca tctctccagt
     6541 ctgggcaaga gggtcctgga gacgctcctg ggtctcagtt ttctaagtgg cctgcggctc
     6601 ctggggctca cgctaacgat gactttgata actggagtac atttcgtcct cgaaccagcg
     6661 ctaatgctag tactatcagt gggagagatg atgacattat gacagagcag gatgacctgg
     6721 gagatgggga cgtgcattcc ctggtgtatc caccctctgc tgccaagatg gcatctacgc
     6781 tgcccagtct gtctgaaatc agcaatccag aaaacatgga gaaccttctg gataatctca
     6841 accttctctc ggctccaaca tctttaactg tgtccaccca gtccgctcct ggcagcatga
     6901 tgcagcaggc accaagcttc ccacacagtg tcaagactac aacacacagc tgggtgtcag
     6961 gaggatctat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     7021 agctggacgg cgacgtaaac ggccacaagt tcagcgtccg cggcgagggc gagggcgatg
     7081 ccaccaacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     7141 ggcccaccct cgtgaccacc ttcggctacg gcgtggcctg cttcagccgc taccccgacc
     7201 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     7261 ccatctcttt caaggacgac ggtacctaca agacccgcgc cgaggtgaag ttcgagggcg
     7321 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     7381 tggggcacaa gctggagtac aacttcaaca gccactacgt ctatatcaca gccgacaagc
     7441 agaagaacag catcaaggct aacttcaaga tccgccacaa cgttgaggac ggcagcgtgc
     7501 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     7561 acaaccacta cctgagccat cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     7621 acatggtcct gctggagttc gtgaccgctg ccggaatcac actgggatcc tctctcgagg
     7681 tctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     7741 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     7801 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     7861 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     7921 ggctctatgg cttctgaggc ggaaagaacc agctgggcct actagatagc ggacccctta
     7981 ccgaaacatc gccgcattct gcagaggagt cgaacttagt gtatgtaaac ttctgaccca
     8041 ctggaattgt gatacagtga attataagtg aaataatctg tctgtaaaca attgttggaa
     8101 aaatgacttg tgtcatgcac aaagtagatg tcctaactga cttgccaaaa ctattgtttg
     8161 ttaacaagaa atttgtggag tagttgaaaa acgagtttta atgactccaa cttaagtgta
     8221 tgtaaacttc cgacttcaac tgtataggga tcc
//
