LOCUS       sequence-219902-        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        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2631..2640
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2637..2639,2640..2642,2643..3356)
                     /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(2637..2639,2640..2642,2643..3353)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2682..2703)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2943..2962)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3290..3311
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3479..3600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3516..3535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3570..3589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3607..4062)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3744..3765)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3956..3975
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4089..4193
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4195..4552
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4195..4386
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4218..4238)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4403..4538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4587..5381
                     /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             4587..5381
                     /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(4641..4660)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5251..5270
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5569..5588)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5613..5660
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5989..6577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6478..6497
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctaga gtcaccatgg
      661 ccgaggcgcc ccaggtggtg gagaccgacc cggacttcga gccgctgccc cggcagcgct
      721 cctgtacctg gccgctgccc aggccggagt ttaaccagtc caactcgacc acctccagtc
      781 cggcgccgtc gggcggcgcg gccgccaacc ccgacgccgc ggcgagcctg gcctcggcgt
      841 ccgctgtcag caccgacttt atgagcaacc tgagcctgct ggaggagagt gaggacttcg
      901 cgcgggcgcc aggctgcgtg gccgtggcgg cggcggctgc ggccagcagg ggcctgtgcg
      961 gggacttcca gggccccgag gcgggctgcg tgcacccagc gccgccacag cccccaccga
     1021 ccgggccgct gtcgcagccc ccacccgtgc ctccctccgc tgccgccgcc gcggggccac
     1081 tcgcgggaca gccgcgcaag accagctcgt cgcgccgcaa cgcgtggggc aacctgtcgt
     1141 acgccgacct catcaccaag gccatcgaga gctcagccga gaagaggctc accctgtcgc
     1201 agatctacga gtggatggtg aagagcgtgc cctacttcaa ggataagggc gacagcaaca
     1261 gctcggcggg ctggaagaat tcaattcgcc acaatctgtc ccttcacagc aagtttattc
     1321 gagtgcagaa tgaaggaact ggaaagagtt cttggtggat gctcaatcca gagggaggca
     1381 agagcggaaa atcaccccgg agaagagctg cgtccatgga caacaacagt aaatttgcta
     1441 agagccgagg gcgggctgct aagaaaaaag catctctcca gtctgggcaa gagggtcctg
     1501 gagacagccc tgggtctcag ttttctaagt ggcctgcgag tcctgggtcc cacagcaacg
     1561 atgactttga taactggagt acatttcgtc ctcgaaccag ctcaaatgct agtaccatca
     1621 gtgggagact ttctcccatc atgacagagc aggatgacct gggagatggg gacgtgcatt
     1681 ccctggtgta tccaccctct gctgccaaga tggcgtctac gctgcccagt ctgtctgaaa
     1741 tcagcaatcc agaaaacatg gagaaccttc tggataatct caaccttctc tcgtccccaa
     1801 catctttaac tgtgtccacc cagtcctcgc ctggcagcat gatgcagcag acaccatgct
     1861 attcgtttgc accgccaaac accagtctaa attcacccag tccaaactac tcaaagtaca
     1921 catacggcca atccagcatg agccctttgc cccagatgcc tatgcagaca cttcaggaca
     1981 gcaaatcaag ttacggagga ttgaaccagt ataactgtgc cccaggactc ttgaaagagt
     2041 tgttgacttc tgactctcct ccccacaatg acattatgtc accggttgat cccggagtgg
     2101 cccaacccaa cagtcgggtc ctgggccaaa atgtaatgat gggccctaat tcggtcatgc
     2161 cagcgtatgg cagccaggca tctcataaca aaatgatgaa ccccagctcc cacacccacc
     2221 ctggacatgc acagcaaacg gcttcggtca acggccgtac cctgccccat gtggtgaaca
     2281 ccatgcctca cacatctgcc atgaaccgct tgacccccgt gaagacacct ttacaagtgc
     2341 ctctgtccca ccccatgcag atgagtgccc tgggcagcta ctcctcggtg agcagctgca
     2401 atggctatgg taggatgggt gtcctccacc aggagaagct cccaagtgac ttggatggca
     2461 tgtttattga gcgcttggac tgtgacatgg agtccatcat tcggaatgac cccatggatg
     2521 gagatacctt ggattttaac tttgataatg tgttgcccaa ccaaagcttc ccacacagtg
     2581 tcaagactac aacacacagc tgggtgtcag gctgcaggga tccaccggtc gccaccatgg
     2641 tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     2701 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     2761 agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     2821 tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc
     2881 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     2941 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     3001 accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc
     3061 tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca
     3121 tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc
     3181 actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc
     3241 tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc
     3301 tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg
     3361 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     3421 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     3481 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     3541 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     3601 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     3661 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     3721 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     3781 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     3841 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     3901 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     3961 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     4021 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     4081 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     4141 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     4201 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     4261 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     4321 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     4381 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     4441 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     4501 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     4561 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4621 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4681 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4741 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     4801 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4861 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4921 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4981 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5041 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5101 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5161 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5221 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5281 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5341 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     5401 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     5461 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     5521 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     5581 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     5641 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     5701 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     5761 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     5821 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5881 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5941 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6001 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6061 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6121 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6181 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6241 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6301 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6361 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6421 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6481 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6541 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6601 acggttcctg gccttttgct ggccttttgc tcacatg
//
