LOCUS       sequence-299701-        6136 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Flag tagged human FOXO3B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6136)
  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 6136)
  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-FOXO3B
FEATURES             Location/Qualifiers
     source          1..6136
                     /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    2188..2322
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2212..2231)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2266..2285
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2435..2455)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2435..2453)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2474..2490)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2474..2490)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2487..2509)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2498..2514
                     /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(2522..2528,2529..2546,2547..2552))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2567..2588
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2705..2722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2876..3464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2956..2975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3635..4426,4427..4495))
                     /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     4258..4277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4496..4600)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4626..5081)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4763..4784)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4975..4994
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5152..6136
                     /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 ctttctccgc 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 ggagccgcac gctggtctcc gggctgctcc ttgaggactc ggtccgggtg
      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 cggacagacg gctcactctg
      661 tcccagatct acgagtggat ggtgagttgc gtgccctact tcaaggataa gggcaacagc
      721 aacagctctg ccggctagaa gaactccatc cggcacaacc tgtcactgca tagtcgattc
      781 atgcgggtcc agaatgaggg aactggcaag agctcttggt ggatcatcaa ccctgatggg
      841 gggaagagcg gaaaagcccc ccggcggcgg gctgtctcca tggacaatag caacaagtat
      901 accaagagcc gtggccgcac agccaagaag aaggcagccc tgcagacagc ccccgaatca
      961 gctgacgaca gtccctccca gctctccaag tggtctggca gccccacgtc acgcagcagt
     1021 gatgagctgg atgcgtggac ggacttccgt tcacgcacca attctaacgc cagcacagtc
     1081 agtggccgcc tgtcgcccat catggcaagc acagagttgg atgaagtcca ggacgacgat
     1141 gcgcctctct cgcccatgct ctacagcagc tcagcgagcc tgtcaccttc agtaagcaag
     1201 ccgtgcacgg tggaactgct acggctgact gacatggcag gcaccatgaa tctgaatgat
     1261 ggggctgact gaaaacctca tggacgacct gctggataac atcacgctcc cgccatccca
     1321 gccatcgccc actgggggac tcatgcagcg gagttctagc ttcccgtata ccaccaaggg
     1381 ctcgggcctg agctccccaa ccagctcctt taacagcacg gtgtttggac cttcatctct
     1441 gaactcccta cgccagtctc ccatgcagac catccaagag aacaagccag ctaccttctc
     1501 ttccatgtca cactatggta accagacact ccaggacctg ctcacttcgg actcacttag
     1561 ccacagcgat gtcatgatga cacagtcgga ccccttgatg tctcaggcca gcaccgctgt
     1621 gtctgcccag aattcccgcc ggaacgtgat gcttcgcaat gatccgatga tgtcctttgc
     1681 tgcccagcct aaccagggaa gtgtggtcaa tcagaacttg ctccaccacc agcaccaaac
     1741 ccagggcgct cttggtggca gccgtgcctt gtcgaattct gtcagcaaca tgggcttgag
     1801 tgagtccagc agccttgggt cagccaaaca ccagcagcag tctcctgtca gccagtctat
     1861 gcaaaccctc tcggactctc tctcaggctc ctccttgtac tcaactagtg caaacctgcc
     1921 cgtcatgggc catgagaagt tccccagcga cttggacctg gacatgttca atgggagctt
     1981 ggaatgtgac atggagtcca ttatccgtag tgaactcatg gatgctgatg ggttggattt
     2041 taactttgat tccctcatct ccacacagaa tgttgttggt ttgaacgtgg ggaacttcac
     2101 tggtgctaag caggcctcat ctcagagctg ggtgccaggc tgaaggatca ctcgagcctc
     2161 tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat tgatgagttt
     2221 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     2281 attgctttat ttgtaaccat tataagctgc aataaacaag ttaacaacaa caattgcatt
     2341 cattttatgt ttcaggttca gggggaggtg tgggaggttt tttaattcgc ggccgcggcg
     2401 ccaatgcatt gggcccggta cccagctttt gttcccttta gtgagggtta attgcgcgct
     2461 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     2521 acaacatacg agccgggagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     2581 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     2641 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     2701 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     2761 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     2821 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     2881 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     2941 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3001 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3061 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3121 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3181 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3241 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3301 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3361 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3421 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     3481 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     3541 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     3601 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     3661 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     3721 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     3781 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     3841 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     3901 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     3961 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4021 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4081 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4141 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4201 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     4261 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     4321 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     4381 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     4441 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     4501 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     4561 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     4621 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     4681 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     4741 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     4801 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     4861 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     4921 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     4981 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     5041 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     5101 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agtcgaccgc
     5161 caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct ggacctgtgc
     5221 caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct ggaccccagc
     5281 caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct ggcactgtgc
     5341 caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct tggccctgtg
     5401 ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt ggctatatgc
     5461 caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct attggccagg
     5521 ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt ttcctattga
     5581 cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt cctaataccg
     5641 cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc ctattgacgt
     5701 catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa
     5761 tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt gacgtcaatg
     5821 ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc aatgacggta
     5881 aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag tacattacta
     5941 ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc ggtaaatggc
     6001 ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa tgacgcaaat
     6061 gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct atataagcaa
     6121 tgctcgttta gggaac
//
