LOCUS       sequence-260832-        6709 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 6709)
  AUTHORS   Qiang L, Banks AS, Accili D
  TITLE     Uncoupling of acetylation from phosphorylation regulates FoxO1 
            function independent of its subcellular localization.
  JOURNAL   J Biol Chem. 2010 Aug 27;285(35):27396-401. Epub 2010 Jun 2.
  PUBMED    20519497
REFERENCE   2  (bases 1 to 6709)
  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."; date: "2010-08-27"; volume: "285(35)"; pages: "27396-401. 
            Epub 2010 Jun 2"
COMMENT     Sequence Label: FoxO1-ADA-GFP
FEATURES             Location/Qualifiers
     source          1..6709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        59..362
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        363..566
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     517..537
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             595..624
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             652..675
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      2647..2656
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2653..2655,2656..2658,2659..3372)
                     /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(2653..2655,2656..2658,2659..3369)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2698..2719)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2959..2978)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3306..3327
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3495..3616
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3532..3551)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3586..3605
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3623..4078)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3760..3781)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3972..3991
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4105..4209
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4211..4568
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4211..4402
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4234..4254)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4419..4554
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4603..5397
                     /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             4603..5397
                     /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(4657..4676)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5267..5286
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5585..5604)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5629..5676
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6005..6593
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6494..6513
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
       61 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      121 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      181 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      241 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      301 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      361 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      421 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      481 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      541 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaattgc catggagcag
      601 aagctgatca gcgaggagga cctgaacgaa ttcagatctg gtaccgccat ggactacaag
      661 gacgacgatg acaaggccga ggcgccccag gtggtggaga ccgacccgga cttcgggccg
      721 ctgccccggc agcgctcctg tgcctggccg ctgcccaggc cggagtttaa ccagtccaac
      781 tcgaccacct ccagtccggc gccgtcgggc ggcgcggccg ccaaccccga cgccgcggcg
      841 agcctggcct cggcgtccgc tgtcagcacc gactttatga gcaacctgag cctgctggag
      901 gagagtgagg acttcgcgcg ggcgccaggc tgcgtggccg tggcggcggc ggctgcggcc
      961 agcaggggcc tgtgcgggga cttccagggc cccgaggcgg gctgcgtgca cccagcgccg
     1021 ccacagcccc caccgaccgg gccgctgtcg cagcccccac ccgtgcctcc ctccgctgcc
     1081 gccgccgcgg ggccactcgc gggacagccg cgcaagacca gctcgtcgcg ccgcaacgcg
     1141 tggggcaacc tgtcgtacgc cgacctcatc accaaggcca tcgagagctc agccgagaag
     1201 aggctcaccc tgtcgcagat ctacgagtgg atggtgaaga gcgtgcccta cttcaaggat
     1261 aagggcgaca gcaacagctc ggcgggctgg aagaattcaa ttcgccacaa tctgtccctt
     1321 cacagcaggt ttattcgagt gcagaatgaa ggaactggaa agagttcttg gtggatgctc
     1381 aatccagagg gaggcaagag cggaaaatca ccccggagaa gagctgcgga catggacaac
     1441 aacagtaaat ttgctaagag ccgagggcgg gctgctaaga aaaaagcatc tctccagtct
     1501 gggcaagagg gtcctagaga cagccctggg tctcagtttt ctaagtggcc tgcgagtcct
     1561 gggtcccaca gcaacgatga ctttgataac tggagtacat ttcgtcctcg aaccagcgca
     1621 aatgctagta ccatcagtgg gagactttct cccatcatga cagagcagga tgacctggga
     1681 gatggggacg tgcattccct ggtgtatcca ccctctgctg ccaagatggc gtctacgctg
     1741 cccagtctgt ctgaaatcag caatccagaa aacatggaga accttctgga taatctcaac
     1801 cttctctcgt ccccaacatc tttaactgtg tccacccagt cctcgcctgg cagcatgatg
     1861 cagcagacac catgctattc gtttgcaccg ccaaacacca gtctaaattc acccagtcca
     1921 aactactcaa agtacacata cggccaatcc agcatgagcc ctttgcccca gatgcctatg
     1981 cagacacttc aggacagcaa atcaagttac ggaggattga accagtataa ctgtgcccca
     2041 ggactcttga aagagttgtt gacttctgac tctcctcccc acaatgacat tatgtcaccg
     2101 gttgatcccg gagtggccca acccaacagt cgggtcctgg gccaaaatgt aatgatgggc
     2161 cctaattcgg tcatgccagc gtatggcagc caggcatctc ataacaaaat gatgaacccc
     2221 agctcccaca cccaccctgg acatgcacag caaacggctt cggtcaacgg ccgtaccctg
     2281 ccccatgtgg tgaacaccat gcctcacaca tctgccatga accgcttgac ccccgtgaag
     2341 acacctttac aagtgcctct gtcccacccc atgcagatga gtgccctggg cagctactcc
     2401 tcggtgagca gctgcaatgg ctatggtagg atgggtgtcc tccaccagga gaagctccca
     2461 agtgacttgg atggcatgtt tattgagcgc ttggactgtg acatggagtc catcattcgg
     2521 aatgacccca tggatggaga taccttggat tttaactttg ataatgtgtt gcccaaccaa
     2581 agcttcccac acagtgtcaa gactacaaca cacagctggg tgtcaggctg cagggatcca
     2641 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     2701 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     2761 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     2821 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
     2881 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     2941 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     3001 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     3061 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     3121 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     3181 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     3241 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     3301 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     3361 ctgtacaagt aaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     3421 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     3481 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3541 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3601 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     3661 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     3721 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     3781 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     3841 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     3901 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     3961 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     4021 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     4081 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4141 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4201 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     4261 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     4321 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4381 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     4441 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     4501 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     4561 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     4621 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     4681 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     4741 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     4801 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     4861 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     4921 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     4981 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     5041 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     5101 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     5161 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     5221 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     5281 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     5341 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     5401 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5461 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5521 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     5581 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     5641 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     5701 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     5761 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     5821 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     5881 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     5941 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6001 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6061 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6121 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6181 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6241 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6301 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6361 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6421 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6481 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6541 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6601 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6661 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcatta
//
