LOCUS       sequence-299549-        5643 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXO4 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5643)
  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 5643)
  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-FOXO4
FEATURES             Location/Qualifiers
     source          1..5643
                     /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    1695..1829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1719..1738)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1773..1792
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1942..1962)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1942..1960)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1981..1997)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1981..1997)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1994..2016)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2005..2021
                     /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(2029..2035,2036..2053,2054..2059))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2074..2095
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2212..2229)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2383..2971)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2463..2482)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3142..3933,3934..4002))
                     /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     3765..3784
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4003..4107)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4133..4588)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4270..4291)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4482..4501
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4659..5643
                     /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 tcccatcgat tcgatccggg gaatgagaat tcagccacag aggctgccgc gatcatagac
      181 ctagatcccg acttcgaacc ccagagccgt ccccgctcct gcacctggcc ccttccccga
      241 ccagagatcg ctaaccagcc gtccgagccg cccgaggtgg agccagatct gggggaaaag
      301 gtacacacgg aggggcgctc agagccgatc ctgttgccct ctcggctccc agagccggcc
      361 gggggccccc agcccggaat cctgggggct gtaacaggtc ctcggaaggg aggctcccgc
      421 cggaatgcct ggggaaatca gtcatatgca gaactcatca gccaggccat tgaaagcgcc
      481 ccggagaagc gactgacact tgcccagatc tacgagtgga tggtccgtac tgtaccctac
      541 ttcaaggaca agggtgacag caacagctca gcaggatgga agaactcgat ccgccacaac
      601 ctgtccctgc acagcaagtt catcaaggtt cacaacgagg ccaccggcaa aagctcttgg
      661 tggatgctga accctgaggg aggcaagagc ggcaaagccc cccgccgccg ggccgcctcc
      721 atggatagca gcagcaagct gctccggggc cgcagtaaag cccccaagaa gaaaccatct
      781 gtgctgccag ctccacccga aggtgccact ccaacgagcc ctgtcggcca ctttgccaag
      841 tggtcaggca gcccttgctc tcgaaaccgt gaagaagccg atatgtggac caccttccgt
      901 ccacgaagca gttcaaatgc cagcagtgtc agcacccggc tgtccccctt gaggccagag
      961 tctgaggtgc tggcggagga aataccagct tcagtcagca gttatgcagg gggtgtccct
     1021 cccaccctca atgaaggtct agagctgtta gatgggctca atctcacctc ttcccattcc
     1081 ctgctatctc ggagtggtct ctctggcttc tctttgcagc atcctggggt taccggcccc
     1141 ttacacacct acagcagctc ccttttcagc ccagcagagg ggcccctgtc agcaggagaa
     1201 gggtgcttct ccagctccca ggctctggag gccctgctca cctctgatac gccaccaccc
     1261 cctgctgacg tcctcatgac ccaggtagat cccattctgt cccaggctcc gactcttctg
     1321 ttgctggggg ggcttccttc ctccagtaag ctggccacgg gcgtcggcct gtgtcccaag
     1381 cccctagagg ctccaggccc cagcagtctg gttcccaccc tttctatgat agcaccacct
     1441 ccagtcatgg caagtgcccc catccccaag gctctgggga ctcctgtgct cacaccccct
     1501 actgaagctg caagccaaga cagaatgcct caggatctag atcttgatat gtatatggag
     1561 aacctggagt gtgacatgga taacatcatc agtgacctca tggatgaggg cgagggactg
     1621 gacttcaact ttgagccaga tccctgagtc atgcctgctc gagcctctag aactatagtg
     1681 agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga caaaccacaa
     1741 ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt gctttatttg
     1801 taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat tttatgtttc
     1861 aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca atgcattggg
     1921 cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg cgtaatcatg
     1981 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     2041 cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     2101 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     2161 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     2221 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     2281 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2341 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2401 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2461 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2521 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2581 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2641 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2701 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2761 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2821 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2881 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2941 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3001 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3061 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3121 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3181 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     3241 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     3301 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     3361 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     3421 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     3481 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     3541 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     3601 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     3661 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     3721 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     3781 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     3841 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     3901 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     3961 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     4021 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     4081 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctaaattgta
     4141 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     4201 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     4261 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     4321 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     4381 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     4441 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     4501 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     4561 gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg caactgttgg
     4621 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa ttcaatatgg
     4681 cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa ttcaatatgg
     4741 cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa ttcaatatgg
     4801 cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa ctggggaggg
     4861 gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca agtccgccat
     4921 attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag gatcaatata
     4981 taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc aatactatgt
     5041 attggcccta tgccatatag tattccatat atgggttttc ctattgacgt agatagcccc
     5101 tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc atagccactc
     5161 ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat atgggcggtc
     5221 ctattgacgt atatggcgcc tcccccattg acgtcaatta cggtaaatgg cccgcctggc
     5281 tcaatgccca ttgacgtcaa taggaccacc caccattgac gtcaatggga tggctcattg
     5341 cccattcata tccgttctca cgccccctat tgacgtcaat gacggtaaat ggcccacttg
     5401 gcagtacatc aatatctatt aatagtaact tggcaagtac attactattg gaagtacgcc
     5461 agggtacatt ggcagtactc ccattgacgt caatggcggt aaatggcccg cgatggctgc
     5521 caagtacatc cccattgacg tcaatgggga ggggcaatga cgcaaatggg cgttccattg
     5581 acgtaaatgg gcggtaggcg tgcctaatgg gaggtctata taagcaatgc tcgtttaggg
     5641 aac
//
