LOCUS       sequence-205984-        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_10A_3D
FEATURES             Location/Qualifiers
     source          1..8250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        171..1353
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          402..1344
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1301..1321
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     complement(2683..2704)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     3291..3312
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3366..3383)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3372..3596
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        4028..4132
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4133..4201,4202..4993)
                     /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(4351..4370)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5164..5752
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5653..5672
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5906..5923
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6297..6316
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    6315..6370
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     6416..6435
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(6535..7134)
                     /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(6618..6638)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7115..7134
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     CDS             complement(7135..7191)
                     /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(7503..7522)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     7772..7790
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     CDS             complement(7930..7950)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 ttcacacgct ctgtccgcca tgtttgtgag tggaagcgcc attacccctt caagcgactg
       61 aaggctgcag ggcctctggt ggcccgcatg gggagaccag acccgccagg cccgcctttc
      121 cgcactcagt ccgggcttac tttattttgt gagacagggt ctcgcctaga ggctccggtg
      181 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      241 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      301 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      361 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      421 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacgc
      481 ccctggctgc agtacgtgat tcttgatccc gagcttcggg ttggaagtgg gtgggagagt
      541 tcgaggcctt gcgcttaagg agccccttcg cctcgtgctt gagttgaggc ctggcttggg
      601 cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg cctgtctcgc tgctttcgat
      661 aagtctctag ccatttaaaa tttttgatga cctgctgcga cgcttttttt ctggcaagat
      721 agtcttgtaa atgcgggcca agatctgcac actggtattt cggtttttgg ggccgcgggc
      781 ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga ggcggggcct gcgagcgcgg
      841 ccaccgagaa tcggacgggg gtagtctcaa gctggccggc ctgctctggt gcctggcctc
      901 gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg cccggtcggc accagttgcg
      961 tgagcggaaa gatggccgct tcccggccct gctgcaggga gctcaaaatg gaggacgcgg
     1021 cgctcgggag agcgggcggg tgagtcaccc acacaaagga aaagggcctt tccgtcctca
     1081 gccgtcgctt catgtgactc cacggagtac cgggcgccgt ccaggcacct cgattagttc
     1141 tcgagctttt ggagtacgtc gtctttaggt tggggggagg ggttttatgc gatggagttt
     1201 ccccacactg agtgggtgga gactgaagtt aggccagctt ggcacttgat gtaattctcc
     1261 ttggaatttg ccctttttga gtttggatct tggttcattc tcaagcctca gacagtggtt
     1321 caaagttttt ttcttccatt tcaggtgtcg tgaggaattt cgacatttaa atttaattaa
     1381 accatggcag aggctcccca ggtggtggag accgacccgg acttcgagcc gctgccccgg
     1441 cagcgctcct gtacctggcc gctgcccagg ccggagttta accagtccaa ctcgaccacc
     1501 tccagtccgg cgccgtcggg cggcgcggcc gccaaccccg acgccgcggc gagcctggcc
     1561 tcggcgtccg ctgtcagcac cgactttatg agcaacctga gcctgctgga ggagagtgag
     1621 gacttcgcgc gggcgccagg ctgcgtggcc gtggcggcgg cggctgcggc cagtaggggc
     1681 ctgtgcgggg acttccaggg ccccgaggcg ggctgcgtgc acccagcgcc gccacagccc
     1741 ccaccgaccg ggccgctgtc gcagccccca cccgtgcctc cctccgctgc cgccgccgcg
     1801 gggccactcg cgggacagcc gcgcaagacc agctcgtcgc gccgcaacgc gtggggcaac
     1861 ctgtcgtacg ccgacctcat caccaaggcc atcgagagct cagccgagaa gaggctcacc
     1921 ctgtcgcaga tctacgagtg gatggtgaag agcgtgccct acttcaagga taagggcgac
     1981 agcaacagct cggcgggctg gaagaatgca attcgccgca atctagccct tcacagcaag
     2041 tttattcgag tgcagaatga aggaactgga aagagttctt ggtggatgct caatccagag
     2101 ggaggcaaga gcggaaaagc tccccggaga agagctgcgt ccatggacaa caacagtaaa
     2161 tttgctaaga gccgagggcg ggctgctaag aaaaaagcat ctctccagtc tgggcaagag
     2221 ggtcctgggg acgctcctgg gtctcagttt tctaagtggc ctgcggctcc tggggctcac
     2281 gctaacgatg actttgataa ctggagtaca tttcgtcctc gaaccagctc aaatgctagt
     2341 actatcagtg ggagagatga tgacattatg acagagcagg atgacctggg agatggggac
     2401 gtgcattccc tggtgtatcc accctctgct gccaagatgg cgtctacgct gcccagtctg
     2461 tctgaaatca gcaatccaga aaacatggag aaccttctgg ataatctcaa ccttctctcg
     2521 gctccaacat ctttaactgt gtccacccag tccgctcctg gcagcatgat gcagcaggca
     2581 ccaagcttcc cacacagtgt caagactaca acacacagct gggtgtcagg aggatctatg
     2641 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2701 gacgtaaacg gccacaagtt cagcgtccgc ggcgagggcg agggcgatgc caccaacggc
     2761 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2821 gtgaccacct tcggctacgg cgtggcctgc ttcagccgct accccgacca catgaagcag
     2881 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catctctttc
     2941 aaggacgacg gtacctacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg
     3001 aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     3061 ctggagtaca acttcaacag ccactacgtc tatatcacag ccgacaagca gaagaacagc
     3121 atcaaggcta acttcaagat ccgccacaac gttgaggacg gcagcgtgca gctcgccgac
     3181 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3241 ctgagccatc agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3301 ctggagttcg tgaccgctgc cggaatcaca ctgggatccc tcgaggtcta gagctcgctg
     3361 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3421 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3481 atcgcattgt ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa
     3541 gggggaggat tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc
     3601 tgaggcggaa agaaccagct gggcctacta gatagcggac cccttaccga aacatcgccg
     3661 cattctgcag aggagtcgaa cttagtgtat gtaaacttct gacccactgg aattgtgata
     3721 cagtgaatta taagtgaaat aatctgtctg taaacaattg ttggaaaaat gacttgtgtc
     3781 atgcacaaag tagatgtcct aactgacttg ccaaaactat tgtttgttaa caagaaattt
     3841 gtggagtagt tgaaaaacga gttttaatga ctccaactta agtgtatgta aacttccgac
     3901 ttcaactgta tagggatcca attttaacaa aatattaacg cttacaattt cctgatgcgg
     3961 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tcaggtggca cttttcgggg
     4021 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     4081 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     4141 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4201 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     4261 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4321 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4381 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4441 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4501 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     4561 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4621 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4681 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     4741 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     4801 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     4861 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     4921 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     4981 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     5041 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5101 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5161 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5221 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5281 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     5341 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5401 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5461 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5521 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5581 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5641 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5701 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5761 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5821 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     5881 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg atccctatac
     5941 agttgaagtc ggaagtttac atacacctta gccaaataca tttaaactca ctttttcaca
     6001 attcctgaca tttaatccta gtaaaaattc cctgtcttag gtcagttagg atcaccactt
     6061 tattttaaga atgtgaaata tcagaataat agtagagaga atgattcatt tcagctttta
     6121 tttctttcat cacattccca gtgggtcaga agtttacata cactcgactc ctctgcagaa
     6181 tgcggcgatg tttcggtaag gggtcccgag tagctagttc atggcagcca gcatatggca
     6241 tatgttgcca aactctaaac caaatactca ttctgatgtt ttaaatgatt tgccctccca
     6301 tatgtccttc cgagtgagag acacaaaaaa ttccaacaca ctattgcaat gaaaataaat
     6361 ttcctttatt agccagaagt cagatgctca aggggcttca tgatgtcccc ataatttttg
     6421 gcagagggaa aaagatctca gtggtatttg tgagccaggg cattggccac accagccacc
     6481 accttctgat aggcagcctg cacctgagga gtgaattcac gacaggcctt cgaatcaggc
     6541 accgggcttg cgggtcatgc accaggtgcg cggtccttcg ggcacctcga cgtcggcggt
     6601 gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg ggcgcggagg tctccaggaa
     6661 ggcgggcacc ccggcgcgct cggccgcctc cactccgggg agcacgacgg cgctgcccag
     6721 acccttgccc tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg cgggctcctt
     6781 gggccggtgc ggcgccagga ggccttccat ctgttgctgc gcggccagcc gggaaccgct
     6841 caactcggcc atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga cgctctccgg
     6901 cgtggtccag accgccaccg cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag
     6961 cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt ggcggtccgg
     7021 atcgacggtg tggcgcgtgg cggggtagtc ggcgaacgcg gcggcgaggg tgcgtacggc
     7081 cctggggacg tcgtcgcggg tggcgaggcg caccgtgggc ttgtactcgg tcatggggcc
     7141 aggattctcc tcgacgtcac cagcctgctt cagcaggctg aagttagtag ctccactgcc
     7201 cttgtacaga tccagtttcg cacgcttagc tgcaggctcg tccatgccgc cggtggagtg
     7261 gcggccctcg gcgcgttcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
     7321 gtccaacttg atgtcgacgt tgtaggcgcc gggcagctgc acgggcttct tggccttgta
     7381 ggtggtcttg acctcagcgt cgtagtggcc gccgtccttc agcttcagcc tctgcttgat
     7441 ctcgcccttc agggcgccgt cctcggggta catccgctcg gaggaggcct cccagcccat
     7501 ggtcttcttc tgcattacgg ggccgtcgga ggggaagttg gtgccgcgca gcttcacctt
     7561 gtagatgaac tcgccgtcct gcagggagga gtcctgggtc acggtcacca cgccgccgtc
     7621 ctcgaagttc atcacgcgct cccacttgaa gcccccgggg aaggacagct tcaagtagtc
     7681 ggggatgtcg gcggggtgct tcacgtaggc cttggagccg tacatgaact gaggggacag
     7741 gatgtcccag gcgaagggca gggggccacc cttggtcacc ttcagcttgg cggtctgggt
     7801 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcacgga
     7861 gccctccatg tgcaccttga agcgcatgaa ctccttgatg atggccatgt tatcctcctc
     7921 gcccttgctc accttccgct ttttcttagg actcatggtg gcactagtgt ctacggctat
     7981 ataccttgaa acaggggcgg cagaagtgac gcaacgcggt tgcgcggacg gcttcctgtg
     8041 ttttcattgg tcgaagcgtt ctgcgcgtgc gcagagattc cccagggaag gcgggccctc
     8101 ttccgaatgt ccaatccgga cattcttccg gttggacctc cgcggtcagc gctgtgcccg
     8161 gcgagagatc acgtggggcg cggaggcggt gctgctgggg cacggccgtc cagcctcggc
     8221 ggccatattt ttgaggggct gttcatctcg
//
