LOCUS       sequence-58130-e        6637 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6637)
  AUTHORS   Frescas D, Valenti L, Accili D
  TITLE     Nuclear trapping of the forkhead transcription factor FoxO1 via 
            Sirt-dependent deacetylation promotes expression of glucogenetic 
            genes.
  JOURNAL   J Biol Chem. 2005 May 27. 280(21):20589-95.
  PUBMED    15788402
REFERENCE   2  (bases 1 to 6637)
  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"; journalName: "J Biol 
            Chem. 2005 May 27. 280(21):20589-95."
COMMENT     Sequence Label: GFP-Foxo1
FEATURES             Location/Qualifiers
     source          1..6637
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2577..2586
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2583..2585,2586..2588,2589..3302)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2583..2585,2586..2588,2589..3299)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2628..2649)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2889..2908)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3236..3257
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3425..3546
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3462..3481)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3516..3535
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3553..4008)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3690..3711)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3902..3921
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4035..4139
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4141..4498
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4141..4332
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4164..4184)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4349..4484
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4533..5327
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4533..5327
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4587..4606)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5197..5216
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5515..5534)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5559..5606
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5935..6523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6424..6443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagagtca
      601 ccatggccga ggcgccccag gtggtggaga ccgacccgga cttcgagccg ctgccccggc
      661 agcgctcctg tacctggccg ctgcccaggc cggagtttaa ccagtccaac tcgaccacct
      721 ccagtccggc gccgtcgggc ggcgcggccg ccaaccccga cgccgcggcg agcctggcct
      781 cggcgtccgc tgtcagcacc gactttatga gcaacctgag cctgctggag gagagtgagg
      841 acttcgcgcg ggcgccaggc tgcgtggccg tggcggcggc ggctgcggcc agcaggggcc
      901 tgtgcgggga cttccagggc cccgaggcgg gctgcgtgca cccagcgccg ccacagcccc
      961 caccgaccgg gccgctgtcg cagcccccac ccgtgcctcc ctccgctgcc gccgccgcgg
     1021 ggccactcgc gggacagccg cgcaagacca gctcgtcgcg ccgcaacgcg tggggcaacc
     1081 tgtcgtacgc cgacctcatc accaaggcca tcgagagctc agccgagaag aggctcaccc
     1141 tgtcgcagat ctacgagtgg atggtgaaga gcgtgcccta cttcaaggat aagggcgaca
     1201 gcaacagctc ggcgggctgg aagaattcaa ttcgccacaa tctgtccctt cacagcaagt
     1261 ttattcgagt gcagaatgaa ggaactggaa agagttcttg gtggatgctc aatccagagg
     1321 gaggcaagag cggaaaatca ccccggagaa gagctgcgtc catggacaac aacagtaaat
     1381 ttgctaagag ccgagggcgg gctgctaaga aaaaagcatc tctccagtct gggcaagagg
     1441 gtcctggaga cagccctggg tctcagtttt ctaagtggcc tgcgagtcct gggtcccaca
     1501 gcaacgatga ctttgataac tggagtacat ttcgtcctcg aaccagctca aatgctagta
     1561 ccatcagtgg gagactttct cccatcatga cagagcagga tgacctggga gatggggacg
     1621 tgcattccct ggtgtatcca ccctctgctg ccaagatggc gtctacgctg cccagtctgt
     1681 ctgaaatcag caatccagaa aacatggaga accttctgga taatctcaac cttctctcgt
     1741 ccccaacatc tttaactgtg tccacccagt cctcgcctgg cagcatgatg cagcagacac
     1801 catgctattc gtttgcaccg ccaaacacca gtctaaattc acccagtcca aactactcaa
     1861 agtacacata cggccaatcc agcatgagcc ctttgcccca gatgcctatg cagacacttc
     1921 aggacagcaa atcaagttac ggaggattga accagtataa ctgtgcccca ggactcttga
     1981 aagagttgtt gacttctgac tctcctcccc acaatgacat tatgtcaccg gttgatcccg
     2041 gagtggccca acccaacagt cgggtcctgg gccaaaatgt aatgatgggc cctaattcgg
     2101 tcatgccagc gtatggcagc caggcatctc ataacaaaat gatgaacccc agctcccaca
     2161 cccaccctgg acatgcacag caaacggctt cggtcaacgg ccgtaccctg ccccatgtgg
     2221 tgaacaccat gcctcacaca tctgccatga accgcttgac ccccgtgaag acacctttac
     2281 aagtgcctct gtcccacccc atgcagatga gtgccctggg cagctactcc tcggtgagca
     2341 gctgcaatgg ctatggtagg atgggtgtcc tccaccagga gaagctccca agtgacttgg
     2401 atggcatgtt tattgagcgc ttggactgtg acatggagtc catcattcgg aatgacccca
     2461 tggatggaga taccttggat tttaactttg ataatgtgtt gcccaaccaa agcttcccac
     2521 acagtgtcaa gactacaaca cacagctggg tgtcaggctg cagggatcca ccggtcgcca
     2581 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     2641 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     2701 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
     2761 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
     2821 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     2881 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     2941 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     3001 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     3061 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     3121 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     3181 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     3241 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     3301 aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3361 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3421 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3481 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3541 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3601 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3661 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3721 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3781 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3841 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3901 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     3961 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4021 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4081 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4141 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4201 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4261 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4321 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4381 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4441 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4501 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4561 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4621 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4681 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4741 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4801 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4861 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4921 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     4981 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5041 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5101 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5161 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5221 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5281 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5341 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5401 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5461 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5521 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5581 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     5641 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     5701 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     5761 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     5821 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     5881 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     5941 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     6001 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     6061 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     6121 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     6181 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     6241 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     6301 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     6361 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     6421 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     6481 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     6541 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     6601 ccctgattct gtggataacc gtattaccgc catgcat
//
