LOCUS       sequence-299548-        6135 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXO3 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6135)
  AUTHORS   Moparthi L, Koch S
  TITLE     A uniform expression library for the exploration of FOX 
            transcription factor biology.
  JOURNAL   Differentiation. 2020 Sep - Oct;115:30-36. doi: 
            10.1016/j.diff.2020.08.002. Epub 2020 Aug 20.
  PUBMED    32858261
REFERENCE   2  (bases 1 to 6135)
  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: 
            "Differentiation. 2020 Sep - Oct"; volume: "115"; pages: "30-36. 
            doi: 10.1016/j.diff.2020.08.002. Epub 2020 Aug 20"
COMMENT     Sequence Label: Flag-FOXO3
FEATURES             Location/Qualifiers
     source          1..6135
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      85..94
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             94..117
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2187..2321
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2211..2230)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2265..2284
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2434..2454)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2434..2452)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2473..2489)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2473..2489)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2486..2508)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2497..2513
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(2521..2527,2528..2545,2546..2551))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2566..2587
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2704..2721)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2875..3463)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2955..2974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3634..4425,4426..4494))
                     /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     4257..4276
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4495..4599)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4625..5080)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4762..4783)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4974..4993
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5151..6135
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcagccacc atggattaca aggacgacga cgataaggga
      121 tccgcagagg caccggcttc cccggccccg ctctctccgc tcgaagtgga gctggacccg
      181 gagttcgagc cccagagccg tccgcgatcc tgtacgtggc ccctgcaaag gccggagctc
      241 caagcgagcc ctgccaagcc ctcgggggag acggccgccg actccatgat ccccgaggag
      301 gaggacgatg aagacgacga ggacggcggg ggacgggccg gctcggccat ggcgatcggc
      361 ggcggcggcg ggagcggcac gctgggctcc gggctgctcc ttgaggactc ggcccgggtg
      421 ctggcacccg gagggcaaga ccccgggtct gggccagcca ccgcggcggg cgggctgagc
      481 gggggtacac aggcgctgct gcagcctcag caaccgctgc caccgccgca gccgggggcg
      541 gctgggggct ccgggcagcc gaggaaatgt tcgtcgcggc ggaacgcctg gggaaacctg
      601 tcctacgcgg acctgatcac ccgcgccatc gagagctccc cggacaaacg gctcactctg
      661 tcccagatct acgagtggat ggtgcgttgc gtgccctact tcaaggataa gggcgacagc
      721 aacagctctg ccggctggaa gaactccatc cggcacaacc tgtcactgca tagtcgattc
      781 atgcgggtcc agaatgaggg aactggcaag agctcttggt ggatcatcaa ccctgatggg
      841 gggaagagcg gaaaagcccc ccggcggcgg gctgtctcca tggacaatag caacaagtat
      901 accaagagcc gtggccgcgc agccaagaag aaggcagccc tgcagacagc ccccgaatca
      961 gctgacgaca gtccctccca gctctccaag tggcctggca gccccacgtc acgcagcagt
     1021 gatgagctgg atgcgtggac ggacttccgt tcacgcacca attctaacgc cagcacagtc
     1081 agtggccgcc tgtcgcccat catggcaagc acagagttgg atgaagtcca ggacgatgat
     1141 gcgcctctct cgcccatgct ctacagcagc tcagccagcc tgtcaccttc agtaagcaag
     1201 ccgtgcacgg tggaactgcc acggctgact gatatggcag gcaccatgaa tctgaatgat
     1261 gggctgactg aaaacctcat ggacgacctg ctggataaca tcacgctccc gccatcccag
     1321 ccatcgccca ctgggggact catgcagcgg agctctagct tcccgtatac caccaagggc
     1381 tcgggcctgg gctccccaac cagctccttt aacagcacgg tgttcggacc ttcatctctg
     1441 aactccctac gccagtctcc catgcagacc atccaagaga acaagccagc taccttctct
     1501 tccatgtcac actatggtaa ccagacactc caggacctgc tcacttcgga ctcacttagc
     1561 cacagcgatg tcatgatgac acagtcggac cccttgatgt ctcaggccag caccgctgtg
     1621 tctgcccaga attcccgccg gaacgtgatg cttcgcaatg atccgatgat gtcctttgct
     1681 gcccagccta accagggaag tttggtcaat cagaacttgc tccaccacca gcaccaaacc
     1741 cagggcgctc ttggtggcag ccgtgccttg tcgaattctg tcagcaacat gggcttgagt
     1801 gagtccagca gccttgggtc agccaaacac cagcagcagt ctcctgtcag ccagtctatg
     1861 caaaccctct cggactctct ctcaggctcc tccttgtact caactagtgc aaacctgccc
     1921 gtcatgggcc atgagaagtt ccccagcgac ttggacctgg acatgttcaa tgggagcttg
     1981 gaatgtgaca tggagtccat tatccgtagt gaactcatgg atgctgatgg gttggatttt
     2041 aactttgatt ccctcatctc cacacagaat gttgttggtt tgaacgtggg gaacttcact
     2101 ggtgctaagc aggcctcatc tcagagctgg gtgccaggct gaaggatcac tcgagcctct
     2161 agaactatag tgagtcgtat tacgtagatc cagacatgat aagatacatt gatgagtttg
     2221 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2281 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2341 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaattcgcg gccgcggcgc
     2401 caatgcattg ggcccggtac ccagcttttg ttccctttag tgagggttaa ttgcgcgctt
     2461 ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca
     2521 caacatacga gccgggagca taaagtgtaa agcctggggt gcctaatgag tgagctaact
     2581 cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     2641 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc
     2701 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     2761 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     2821 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2881 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2941 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     3001 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     3061 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     3121 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     3181 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     3241 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     3301 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     3361 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     3421 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     3481 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     3541 attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat
     3601 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc
     3661 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat
     3721 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc
     3781 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag
     3841 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag
     3901 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt
     3961 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg
     4021 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt
     4081 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc
     4141 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc
     4201 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa
     4261 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg
     4321 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc
     4381 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag
     4441 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt
     4501 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt
     4561 tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc
     4621 acctaaattg taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc
     4681 tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc
     4741 gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac
     4801 tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca
     4861 ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg
     4921 agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag
     4981 aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc
     5041 accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc attcaggctg
     5101 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gtcgaccgcc
     5161 aattcaatat ggcgtatatg gactcatgcc aattcaatat ggtggatctg gacctgtgcc
     5221 aattcaatat ggcgtatatg gactcgtgcc aattcaatat ggtggatctg gaccccagcc
     5281 aattcaatat ggcggacttg gcaccatgcc aattcaatat ggcggacctg gcactgtgcc
     5341 aactggggag gggtctactt ggcacggtgc caagtttgag gaggggtctt ggccctgtgc
     5401 caagtccgcc atattgaatt ggcatggtgc caataatggc ggccatattg gctatatgcc
     5461 aggatcaata tataggcaat atccaatatg gccctatgcc aatatggcta ttggccaggt
     5521 tcaatactat gtattggccc tatgccatat agtattccat atatgggttt tcctattgac
     5581 gtagatagcc cctcccaatg ggcggtccca tataccatat atggggcttc ctaataccgc
     5641 ccatagccac tcccccattg acgtcaatgg tctctatata tggtctttcc tattgacgtc
     5701 atatgggcgg tcctattgac gtatatggcg cctcccccat tgacgtcaat tacggtaaat
     5761 ggcccgcctg gctcaatgcc cattgacgtc aataggacca cccaccattg acgtcaatgg
     5821 gatggctcat tgcccattca tatccgttct cacgccccct attgacgtca atgacggtaa
     5881 atggcccact tggcagtaca tcaatatcta ttaatagtaa cttggcaagt acattactat
     5941 tggaagtacg ccagggtaca ttggcagtac tcccattgac gtcaatggcg gtaaatggcc
     6001 cgcgatggct gccaagtaca tccccattga cgtcaatggg gaggggcaat gacgcaaatg
     6061 ggcgttccat tgacgtaaat gggcggtagg cgtgcctaat gggaggtcta tataagcaat
     6121 gctcgtttag ggaac
//
